Figure 6: Downstream investigation of HSD11b1 and CEBPb by transient knockdown and principal component analysis.

a) Schematic of the microarray pipeline for combinatorial investigation of the phenotype of HSCs as a function of 3ECMs, 2 stiffness, two transient gene knockdowns and a drug treatment. b) Box Plot of quantification of cellular eccentricity of HSCs on C4TC, treated with DMSO as function for the gene knockdown on each stiffness. c) Representative Image for the cellular morphology quantifications for NC and HSD KD HSCS on 1kPa, C4TC, DMSO. Scale Bar 100 microns. d) Box Plot of quantification of the nuclear area of HSCs on C4TC, treated with DMSO as function for the gene knockdown on each stiffness. e) Representative images of DAPI stain for the nuclear area quantifications for NC and HSD KD HSCS on 1kPa, C4TC, DMSO. Scale Bar 100 microns. f) Correlation plot of measured epigenetic and phenotypic readouts to PC1 and PC2 respectively. c) Principal component plane for 8 readouts as a function of the specific microenvironment (ECM, stiffness, gene knockdown and drug treatment). Every data point on the plane represents a specific microenvironmental condition. Boxplots: ‘*’: p-value <0.05; ‘****’: p-value <0.0001, calculated using Wilcox test in R, n>= 3 biological replicates (independent experiments) and n>= 10 technical replicates (individual islands).