Analysis of transcription factor activity (Affymetrix combo protein–DNA array) for Ctrl, Blebb, and Blebb + AP conditions. Each square corresponds to a specific transcription factor (TF) activity and is color coded to show the relative activity. To identify TFs regulated by nuclear shape, we selected TFs that were more active in Ctrl and Blebb + AP compared to blebbistatin‐treated cells (Blebb).
TF's target genes and GAPDH mRNA levels were analyzed by real‐time‐qPCR. Results are expressed as relative mRNA expression levels. Data are presented as mean ± s.e.m. (n = 4 minimum, t‐test *P < 0.5, **P < 0.05, ***P < 0.01, ****P < 0.001).
TEAD activity was analyzed in HeLa cells co‐transfected with a Renilla plasmid as a luciferase reporter plasmid controlled by the TEAD‐responsive promoter, and with a Renilla plasmid as a gene reporter; HeLa cells were cultured on 1 and 50 kPa (n = 3, t‐test ***P < 0.01) or treated with blebbistatin and blebbistatin + AP (n = 6; t‐test *P < 0.5). Data are presented as mean ± s.e.m.
AP1 activity was assessed in the indicated conditions using AP1 reporter assay. Data are presented as mean ± s.e.m. (n = 5, t‐test ****P < 0.001).
Heatmap of mRNA expression profiles of genes regulated by TEAD and AP1.
