Figure 4.

TSC22D1 acts downstream of C/EBPβ on the inflammatory secretome. (A) Cell proliferation assay of HDF (Tig3(et)-16i) expressing the indicated constructs. Cells were fixed and stained 10 days after BRAF^E600 expression. (B, C, E–G) Regulation of TSC22D1.2, C/EBPβ, IL8, IL6 and IL1β transcripts of the samples described in (A) as determined by qRT–PCR. Measurements are based on at least three independent experiments and standardized to the BRAF^E600-expressing senescent cells. Error bars indicate s.d. (D) Protein levels of TSC22D1, BRAF and C/EBPβ were analysed by immunoblotting using the samples described in (A) (arrowheads indicate the three TSC22D1.2 proteins (see Figure 1D), open triangles indicate C/EBPβ proteins). β-Actin serves as a loading control. (H, I) Microarray gene-expression data of BRAF^E600-expressing senescent cells were correlated to the expression profile found in cells bypassing BRAF^E600-induced arrest upon depletion of either TSC22D1 or C/EBPβ. (H) Two-sample correlation plot. The Pearson product–moment coefficient, reflecting the degree of linear relationship between two variables, is 0.56. (I) Venn diagram displaying the amount of significantly regulated genes with a P-value <0.02. In all, 47 702 transcripts were not significantly regulated. No anti-regulated genes were found.