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FC_step4_Glut2_v4.m
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FC_step4_Glut2_v4.m
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tic
clear all
close all
addpath('/data/matlab_functions/')
terminate(pyenv)
pyenv("ExecutionMode","OutOfProcess")
savefig_flag = 0;
savefig_pdf = 0;
load afterloading_QC3000_FC_06-Oct-2021
IEG_markers = loadCellFile('IEG_cluster.txt');
[~,in_ieg] = ismember(IEG_markers,geneid);
tot_mol = sum(data);
tot_mol(tot_mol>3e4) = 3e4;
tot_genes = sum(data>0);
cellidclsuter_gaba = loadCellFile('cellidCluster_step3_Glut2_FC_24-Oct-2021.txt');
% cellidclsuter_gaba = [cellidclsuter_gaba, cellfun(@(x,y) ['gaba_',num2str(x),'_',num2str(y)],cellidclsuter_gaba(:,2),cellidclsuter_gaba(:,3),'UniformOutput',0)];
gaba_clustername = loadCellFile('ClusterOrder_step3_Glut2_FC_24-Oct-2021.txt');
cellidclsuter_gaba = [cellidclsuter_gaba,cell(length(cellidclsuter_gaba),1)];
c = cell2mat(cellidclsuter_gaba(:,2));
for i=1:length(gaba_clustername)
ind = find(c==gaba_clustername{i});
cellidclsuter_gaba(ind,end) = repmat(gaba_clustername(i,2),length(ind),1);
end
cellid_clusters = cell(length(cellid),4);
[~,loc]= ismember(cellidclsuter_gaba(:,1),cellid);
cellid_clusters(loc(loc>0),:) = cellidclsuter_gaba;
validcells = cell2mat(cellfun(@(x) ~isempty(x), cellid_clusters(:,1),'UniformOutput',0));
sum(validcells)
data = data(:,validcells);
cellid = cellid(validcells);
cellid_clusters = cellid_clusters(validcells,:);
sample = sample(validcells);
fc_time = fc_time(validcells);
amy_flag = amy_flag(validcells);
batch_flag = batch_flag(validcells);
[~,xi] = sort(cell2mat(cellid_clusters(:,2)));
data = data(:,xi);
cellid = cellid(xi);
cellid_clusters = cellid_clusters(xi,:);
sample = sample(xi);
fc_time = fc_time(xi);
amy_flag = amy_flag(xi);
batch_flag = batch_flag(xi);
sample_uni = {'08-1','23-1', '23-3', '45-3','46-3', '74-1', '74-2',.....
'10-1', '18-1', '19-1', '76-1', '76-2',.....
'42-1', '42-2','46-1', '46-2',.....
'41-1', '41-2', '45-1', '45-2',....
'75-1', '75-2', '75-3'};
data = normalize(data,'norm');
data = round(data./repmat(sum(data),length(data(:,1)),1)*20e3);
[median(sum(data(:,batch_flag==1))),median(sum(data(:,batch_flag==2))),median(sum(data(:,batch_flag==3)))]
[median(sum(data(:,batch_flag==1)>0)),median(sum(data(:,batch_flag==2)>0)),median(sum(data(:,batch_flag==3)>0))]
% % % % % % % % % % % % % % % % % % %
clusteruni = gaba_clustername(:,2);
T_cells_tmp = zeros(length(cellid),1);
clustersize = zeros(length(clusteruni),1);
for i=1:length(clusteruni)
ind = find(strcmpi(cellid_clusters(:,4),clusteruni{i}));
T_cells_tmp(ind) = i;
clustersize(i) = length(ind);
end
T_cells_tmp_uni = unique(T_cells_tmp);
fc_time_uni = unique(fc_time);
% % % % % % % % % % % % % % % % % % %
%%
gn = 'dlk1';
g = find(strcmpi(gn,geneid));
figure;
set(gcf,'color','w','position',[20,20,1200,1000],'Name',gn);
b = 1;
if b==2;
[ha, pos] = tight_subplot(4, ceil(length(unique(T_cells_tmp(:,b)))/4), [0.02,0.02], [0.02,0.02], [0.02,0.02]);
elseif b==1;
[ha, pos] = tight_subplot(8, ceil(length(unique(T_cells_tmp(:,b)))/8), [0.02,0.02], [0.02,0.02], [0.02,0.02]);
end
logflag = 0;
p99 = prctile(data(g,:),99.99);
colorvec = distinguishable_colors(3);
for k=1:length(T_cells_tmp_uni)
axes(ha(k))
c=k;
t_ed = zeros(5,1);
t_av = zeros(5,1);
t_75 = zeros(5,1);
p90 = prctile(data(g,T_cells_tmp(:,b)==c),90);%max(ceil(prctile(data(g,T_cells_tmp(:,b)==c),90)),1);
for i=[1:5]
gr2 = find(T_cells_tmp(:,b)==c & fc_time==fc_time_uni(i));%find(fc_time_sorted==fc_time_uni(i));%
bfgr2 = batch_flag(gr2);
y = (data(g,gr2));
if logflag ==1
y = y+1;
yscale = 'log';
else
yscale = 'linear';
end
if length(y)>10
[f,xi] = ksdensity(y,'support',[0-0.1,0.1+max(y)]);
fi = interp1(xi,f,y);
fi = fi/max(fi);
x1 = i + fi'.*(0.9*rand(length(gr2),1)-0.45);
y1 = 0.5*rand(length(gr2),1)-0.1+y';
for jj=1:3
v = bfgr2==jj;
plot(x1(v),y1(v) ,'.','color',colorvec(jj,:)); hold on;
end
else
plot(i+0.7*rand(length(gr2),1)-0.1,0.5*rand(length(gr2),1)-0.1+data(g,gr2)','.'); hold on;
end
% plot(i+0.5*rand(length(gr2),1)-0.1,0.5*rand(length(gr2),1)-0.1+data(g,gr2)','.'); hold on;
t_ed(i) = median(data(g,gr2));
t_av(i) = mean(data(g,gr2));
t_75(i) = prctile(data(g,gr2),75);
f90(i) = sum(data(g,gr2)>p90);
pp90(i) = binocdf(f90(i),length(gr2),0.1,'upper');
end
plot([0,6],p90*[1,1],'--k');
% plot(t_ed,'-sk'); hold on;
% plot(t_av,'-or'); hold on;
% plot(t_75,'-dg'); hold on;
axis tight
yl = get(gca,'ylim');
if k>=(length(T_cells_tmp_uni)-ceil(length(unique(T_cells_tmp(:,b)))/8))
set(gca,'xtick',[1:5],'XTickLabel',[{'0','2','8','24','recall'}],'yscale','linear','ylim',[-0.5,p99+1],'YScale',yscale)
else
set(gca,'xtick',[1:5],'XTickLabel',cell(1,5),'yscale','linear','ylim',[-0.5,p99+1],'YScale',yscale)
end
text([1:5],p99*0.7*ones(1,5),cellfun(@(x) num2str(x,2), m2c(-log10(pp90)),'UniformOutput',0),'fontsize',6);
title(regexprep(clusteruni{c},'_','-'));%[gn,',c=',num2str(c)])
end
if 1==1
% savefig(gcf,['clustermarkers_violin_final_step3_FC_GABA_',date,'.fig'])
eval(['export_fig ',gn,'_violin_FC_Glut2_',date,'.pdf']);
end
%%
%%
nsim = 100;
t_sample_uni = [0,0,0,0,0,0,0,2,2,2,2,2,8,8,8,8,24,24,24,24,28,28,28];
subset_p = [randi(7,[nsim,6]),randi([8,12],[nsim,4]),randi([13,16],[nsim,3]),randi([17,20],[nsim,3]),randi([21,23],[nsim,2])];
pbino_cell = cell(nsim,1);
generank = zeros(length(geneid),nsim);
for jjj=1:nsim
jjj
s = ismember(sample, sample_uni(subset_p(jjj,:)));
k = 0;
b = 1;
pbino_mat = ones(length(geneid),5*length(T_cells_tmp_uni));
for c1=1:length(unique(T_cells_tmp(:,b)))
c1;
fc_time_uni = [0,2,8,24,28];
p90gene = prctile(data(:,s & T_cells_tmp(:,b)==c1),90,2);
for i=[1,2,3,4]
k = k+1;
gr2 = find(s & T_cells_tmp(:,b)==c1 & fc_time==fc_time_uni(i));
x90 = sum(data(:,gr2)>repmat(p90gene,1,length(gr2)),2);
x90(p90gene==0) = nan;
pbino = binocdf(x90,length(gr2),0.1,'upper');
pbino(isnan(pbino)) = 1;
pbino_mat(:,k) = pbino;
end
end
x = sum(pbino_mat(:,:)<1e-2,2);
[~,xi] = sort(x,'descend');
generank(xi,jjj) = [1:length(xi)]';
pbino_cell{jjj} = pbino_mat;
end
f = sum(generank<300,2) - sum(generank>1000,2);
f = sum(generank,2) ;
f = mean(generank<500,2);
[~,xi] = sort(f,'descend');
[geneid(xi(1:100)),m2c(f(xi(1:100)))];
hitgenes = [geneid(f>0.5),m2c(f(f>0.5))]
saveCellFile([hitgenes],['hitgenes_glut2_',date,'.txt'])
in = find(sum(data>0,2)>100 & sum(data>0,2)<length(data(1,:))*0.9);
k = 0;
b = 1;
pbino_mat = ones(length(geneid),5*length(T_cells_tmp_uni));
pbino_mat_rnd = ones(length(geneid),5*length(T_cells_tmp_uni));
fc_time_rnd = fc_time;
p90_mat = zeros(size(pbino_mat));
for c1=1:length(unique(T_cells_tmp(:,b)))
c1
fc_time_uni = [0,2,8,24,28];
t = fc_time_rnd(T_cells_tmp(:,b)==c1);
fc_time_rnd(T_cells_tmp(:,b)==c1) = t(randperm(length(t)));
p90gene = prctile(data(:,T_cells_tmp(:,b)==c1),90,2);
for i=[1,2,3,4,5]
k = k+1
gr2 = find(T_cells_tmp(:,b)==c1 & fc_time==fc_time_uni(i));
gr2_rnd = find(T_cells_tmp(:,b)==c1 & fc_time_rnd==fc_time_uni(i));
x90 = sum(data(:,gr2)>repmat(p90gene,1,length(gr2)),2);
x90(p90gene==0) = nan;
pbino = binocdf(x90,length(gr2),0.1,'upper');
pbino(isnan(pbino)) = 1;
pbino_mat(:,k) = pbino;
x90_rnd = sum(data(:,gr2_rnd)>repmat(p90gene,1,length(gr2_rnd)),2);
x90_rnd(p90gene==0) = nan;
pbino_rnd = binocdf(x90_rnd,length(gr2_rnd),0.1,'upper');
pbino_rnd(isnan(pbino_rnd)) = 1;
pbino_mat_rnd(:,k) = pbino_rnd;
p90_mat(:,k) = prctile(data(:,gr2),90,2);
end
end
pbino_mat(pbino_mat<1e-300) = 1e-300;
save(['pbino_glut2_',date],'pbino_mat','geneid','clusteruni')
save(['p90mat_glut2_',date],'p90_mat','geneid','clusteruni')
[~,inhit] = ismember(hitgenes(:,1),geneid);
m = -log10(pbino_mat(inhit,:));
z = linkage(m,'ward','correlation');
d = corr_mat(m');
leaforder = optimalleaforder(z,squareform(1-d,'tovector'));
tp_order = reshape([1:5*length(T_cells_tmp_uni)],5,length(T_cells_tmp_uni))';
tp_order = tp_order(:);
figure('position',[200,200,1200,1000],'color','w');
set(gca,'Position',[0.1,0.03,0.85,0.9])
imagesc((m(leaforder,tp_order)),[0,3]); hold on;
colormap('summer');
freezeColors(gca);
yl = get(gca,'ylim');
for i=1:length(fc_time_uni)-1
plot(i*length(T_cells_tmp_uni)*[1,1]+0.5,yl,'-k');
end
set(gca,'ytick',[1:length(leaforder)],'YTickLabel',hitgenes(leaforder,1),'fontsize',10,'xtick'...
,[length(T_cells_tmp_uni)/2:length(T_cells_tmp_uni):5*length(T_cells_tmp_uni)],'XTickLabel',{'0','2','8','24','recall'})
mnew = m(leaforder,tp_order);
c_axis = [];
for i=1:5
ind = (i-1)*length(T_cells_tmp_uni)+1:i*length(T_cells_tmp_uni);
mtmp = mnew(:,ind)';
z = linkage(mtmp,'ward','correlation');
d = corr_mat(mtmp');
leaforder_c = optimalleaforder(z,squareform(1-d,'tovector'));
mnew(:,ind) = mnew(:,ind(leaforder_c));
c_axis = [c_axis,leaforder_c];
end
figure('position',[200,200,1200,1000],'color','w');
set(gca,'Position',[0.1,0.03,0.85,0.9])
imagesc((mnew),[0,3]); hold on;
colormap('summer');
freezeColors(gca);
yl = get(gca,'ylim');
for i=1:length(fc_time_uni)-1
plot(i*length(T_cells_tmp_uni)*[1,1]+0.5,yl,'-k');
end
set(gca,'ytick',[1:length(leaforder)],'YTickLabel',hitgenes(leaforder,1),'fontsize',10,'xtick'...
,[length(T_cells_tmp_uni)/2:length(T_cells_tmp_uni):5*length(T_cells_tmp_uni)],'XTickLabel',{'0','2','8','24','recall'})
%%
in = find(sum(data>0,2)>100 & sum(data>0,2)<length(data(1,:))*0.9);
k = 0;
pbino_mat = ones(length(geneid),5*length(T_cells_tmp_uni));
for c1=1:length(unique(T_cells_tmp(:,b)))
c1
fc_time_uni = [0,2,8,24,28];
p90gene = ((prctile(data(:,T_cells_tmp(:,b)==c1),90,2)));
for i=[1,2,3,4,5]
k = k+1
gr2 = find(T_cells_tmp(:,b)==c1 & fc_time==fc_time_uni(i));
x90 = sum(data(:,gr2)>repmat(p90gene,1,length(gr2)),2);
x90(p90gene==0) = nan;
pbino = binocdf(x90,length(gr2),0.1,'upper');
pbino(isnan(pbino)) = 1;
pbino(pbino<1e-300) = 1e-300;
% [~,ptt] = ttest2(data(in,gr1)',data(in,gr2)','tail','both');
pbino_mat(:,k) = pbino;
end
end
x = sum(pbino_mat(in,:)<1e-2,2);
[~,xi] = sort(x,'descend');
g = [geneid(in(xi(1:100))),m2c(x(xi(1:100)))]
m = -log10(pbino_mat(in(xi(1:100)),:));
m_ieg = -log10(pbino_mat(in_ieg,:));
msum = reshape(sum(m>=2),length(T_cells_tmp_uni),5);
figure('position',[200,200,1200,1000],'color','w');
[ha, pos] = tight_subplot(1, 5, [0.05,0.05], [0.02,0.02], [0.05,0.05]);
for i=1:length(fc_time_uni)
s = msum(:,i);
[ssort,sxi] = sort(s,'descend');
axes(ha(i));
scatter(ssort,[1:length(T_cells_tmp_uni)],clustersize(sxi));
set(ha(i),'ytick',[1:length(T_cells_tmp_uni)],'yticklabel',regexprep(clusteruni(sxi),'_','-'),'fontsize',6)
title(['t=',num2str(fc_time_uni(i))])
end
msum = reshape(sum(m_ieg>1),length(T_cells_tmp_uni),5);
figure('position',[200,200,1200,1000],'color','w');
[ha, pos] = tight_subplot(1, 5, [0.05,0.05], [0.02,0.02], [0.05,0.05]);
for i=1:length(fc_time_uni)
s = msum(:,i);
[ssort,sxi] = sort(s,'descend');
axes(ha(i));
scatter(ssort,[1:length(T_cells_tmp_uni)],clustersize(sxi));
set(ha(i),'ytick',[1:length(T_cells_tmp_uni)],'yticklabel',regexprep(clusteruni(sxi),'_','-'),'fontsize',6)
title(['t=',num2str(fc_time_uni(i))])
end
figure;
set(gcf,'color','w','position',[20,20,1200,1000],'Name',gn);
[ha, pos] = tight_subplot(8, ceil(length(unique(T_cells_tmp(:,1)))/8), [0.02,0.02], [0.02,0.02], [0.02,0.02]);
for k=1:length(T_cells_tmp_uni)
axes(ha(k))
bar(msum(k,:));
title(regexprep(clusteruni(k),'_','-'))
end
z = linkage(m,'ward','correlation');
d = corr_mat(m');
leaforder = optimalleaforder(z,squareform(1-d,'tovector'));
tp_order = reshape([1:5*length(T_cells_tmp_uni)],5,length(T_cells_tmp_uni))';
tp_order = tp_order(:);
figure('position',[200,200,1200,1000],'color','w');
set(gca,'Position',[0.1,0.03,0.85,0.9])
imagesc((m(leaforder,tp_order)),[0,3]); hold on;
colormap('summer');
freezeColors(gca);
yl = get(gca,'ylim');
for i=1:length(fc_time_uni)-1
plot(i*length(T_cells_tmp_uni)*[1,1]+0.5,yl,'-k');
end
set(gca,'ytick',[1:length(leaforder)],'YTickLabel',g(leaforder),'fontsize',10,'xtick'...
,[length(T_cells_tmp_uni)/2:length(T_cells_tmp_uni):5*length(T_cells_tmp_uni)],'XTickLabel',{'0','2','8','24','recall'})
if savefig_flag==100
% savefig(gcf,['clustermarkers_violin_final_step3_FC_GABA_',date,'.fig'])
eval(['export_fig top100_mostdiff_FC_Glut2_',date,'.pdf']);
end
inrmv = (sum(m_ieg,2)==0);
g = IEG_markers(~inrmv);
m_ieg(inrmv,:) = [];
z = linkage(m_ieg,'ward','correlation');
d = corr_mat(m_ieg');
leaforder = optimalleaforder(z,squareform(1-d,'tovector'));
tp_order = reshape([1:5*length(T_cells_tmp_uni)],5,length(T_cells_tmp_uni))';
tp_order = tp_order(:);
figure('position',[200,200,1200,1000],'color','w');
set(gca,'Position',[0.1,0.03,0.85,0.9])
imagesc((m_ieg(leaforder,tp_order)),[0,3]); hold on;
colormap('summer');
freezeColors(gca);
yl = get(gca,'ylim');
for i=1:length(fc_time_uni)-1
plot(i*length(T_cells_tmp_uni)*[1,1]+0.5,yl,'-k');
end
set(gca,'ytick',[1:length(leaforder)],'YTickLabel',g(leaforder),'fontsize',10,'xtick'...
,[length(T_cells_tmp_uni)/2:length(T_cells_tmp_uni):5*length(T_cells_tmp_uni)],'XTickLabel',{'0','2','8','24','recall'})
% for i=1:length(T_cells_tmp_uni)-1
% plot(i*5*[1,1]+0.5,yl,'-k');
% end
% set(gca,'ytick',[1:length(leaforder)],'YTickLabel',g(leaforder),'fontsize',10,'xtick'...
% ,[3:6:5*length(T_cells_tmp_uni)],'XTickLabel',regexprep(clusteruni,'_','-'),'XTickLabelRotation',45)
%%
%%
% [ind_gr_tmp_mark,cells_bor,gr_center] = markertablefeatures(T_cells_tmp(:,1),data_sorted_all,1);
list = {'Lynx1','Diras2','Slit2','Cdh8'....
,'Neurod6','Bmp3','Rspo2','Man1a','Sim1','C1ql1','Tac1','Meis2','Cartpt','Prox1'};
% list = {'Fth1','Ftl1','Scrib','Tfr2','Tfrc','Slc11a2','Slc40a1','Aco1','Ireb2'};
figure;
set(gcf,'color','w','position',[20,20,1000,1000]);
[ha, pos] = tight_subplot(length(list), 1, [0.01,0.01], [0.02,0.01], [0.15,0.01]);
logflag = 0;
for jjj=1:length(list)
gn = list{jjj};
g = find(strcmpi(gn,geneid));
p99 = prctile(data(g,:),100);
axes(ha(jjj))
p90 = max(ceil(prctile(data(g,:),90)),1);
t_ed = zeros(length(T_cells_tmp_uni),1);
t_av = zeros(length(T_cells_tmp_uni),1);
t_75 = zeros(length(T_cells_tmp_uni),1);
for k=1:length(T_cells_tmp_uni)
c=k;
gr2 = find(T_cells_tmp(:,1)==c);
y = (data(g,gr2));
if logflag ==1
y = y+1;
yscale = 'log';
else
yscale = 'linear';
end
if length(y)>10
[f,xi] = ksdensity(y,'support',[0-0.1,0.1+max(y)]);
fi = interp1(xi,f,y);
fi = fi/max(fi);
plot(k + fi'.*(0.9*rand(length(gr2),1)-0.45), 0.5*rand(length(gr2),1)-0.1+y','.'); hold on;
else
plot(k+0.7*rand(length(gr2),1)-0.1,0.5*rand(length(gr2),1)-0.1+data(g,gr2)','.'); hold on;
end
t_ed(k) = median(data(g,gr2));
t_av(k) = mean(data(g,gr2));
t_75(k) = prctile(data(g,gr2),75);
end
axis tight
yl = get(gca,'ylim');
if jjj==length(list)
set(gca,'xtick',[1:length(T_cells_tmp_uni)],'ylim',[-0.5,p99+1],'YScale',yscale);
else
set(gca,'xtick',[1:length(T_cells_tmp_uni)],'xticklabel',cell(size(T_cells_tmp_uni)),'ylim',[-0.5,p99+1],'YScale',yscale)
end
text(-3,p99/2,[gn])
box off
end
if savefig_flag==100
% savefig(gcf,['clustermarkers_violin_final_step3_FC_GABA_',date,'.fig'])
eval(['export_fig markers1_violin_final_step3_FC_Glut2_',date,'.pdf']);
end
%%
% [ind_gr_tmp_mark,cells_bor,gr_center] = markertablefeatures(T_cells_tmp(:,1),data_sorted_all,1);
% list = {'Maf','Nfib','Zeb2','Htr3a','Sncg','Vip','Chat','Reln','Ndnf','Lamp5','Pvalb','Sst','Chodl'};%geneid(ind_gr_tmp_mark(1:end));
list = {'Mpped2','Krt12','Gpr88','Trhr','Cd24a','Igfbpl1','Pde11a','Trh','Rab3b','Kctd1','Gsg1l'};
% list = {'Meis2','Maf','Zeb2','Prlr','Lhx6','Npas1','Igfbp4','Sst','Cryab','Penk','Moxd1','Prkcd','Unc5b','Ucn3','Pthlh','Vip','Chat','Htr3a','Vdr'};
% list = {'Fth1','Ftl1','Scrib','Tfr2','Tfrc','Slc11a2','Slc40a1','Aco1','Ireb2'};
figure;
set(gcf,'color','w','position',[20,20,1000,1000]);
[ha, pos] = tight_subplot(length(list), 1, [0.01,0.01], [0.02,0.01], [0.15,0.01]);
logflag = 0;
for jjj=1:length(list)
gn = list{jjj};
g = find(strcmpi(gn,geneid));
p99 = prctile(data(g,:),100);
axes(ha(jjj))
p90 = max(ceil(prctile(data(g,:),90)),1);
t_ed = zeros(length(T_cells_tmp_uni),1);
t_av = zeros(length(T_cells_tmp_uni),1);
t_75 = zeros(length(T_cells_tmp_uni),1);
for k=1:length(T_cells_tmp_uni)
c=k;
gr2 = find(T_cells_tmp(:,1)==c);
y = (data(g,gr2));
if logflag ==1
y = y+1;
yscale = 'log';
else
yscale = 'linear';
end
if length(y)>10
[f,xi] = ksdensity(y,'support',[0-0.1,0.1+max(y)]);
fi = interp1(xi,f,y);
fi = fi/max(fi);
plot(k + fi'.*(0.9*rand(length(gr2),1)-0.45), 0.5*rand(length(gr2),1)-0.1+y','.'); hold on;
else
plot(k+0.7*rand(length(gr2),1)-0.1,0.5*rand(length(gr2),1)-0.1+data(g,gr2)','.'); hold on;
end
t_ed(k) = median(data(g,gr2));
t_av(k) = mean(data(g,gr2));
t_75(k) = prctile(data(g,gr2),75);
end
axis tight
yl = get(gca,'ylim');
if jjj==length(list)
set(gca,'xtick',[1:length(T_cells_tmp_uni)],'ylim',[-0.5,p99+1],'YScale',yscale);
else
set(gca,'xtick',[1:length(T_cells_tmp_uni)],'xticklabel',cell(size(T_cells_tmp_uni)),'ylim',[-0.5,p99+1],'YScale',yscale)
end
text(-3,p99/2,[gn])
box off
end
if savefig_flag==100
% savefig(gcf,['clustermarkers_violin_final_step3_FC_GABA_',date,'.fig'])
eval(['export_fig markers2_violin_final_step3_FC_Glut2_',date,'.pdf']);
end
%%