Fig. 3

TNF-induced oncogenic signaling pathways were interrupted upon CSMD1 overexpression in glioma cells. Ctrl and CSMD1 H4, U-118 and U-87 cells were serum-starved for 2 h followed by treatment with TNF at a concentration of 25 ng/ml for 1, 2 and 4 h. A negative control was established by treating cells with BSA. A&E Representative western blots of glioma cell lines treated with BSA or TNF for the indicated time points immunodetecting pP65-Ser536, total P65 and β-actin. β-actin was used as an internal control. B Densitometry analysis of pP65-Ser536/β-actin, pP65-Ser536/Total P65 and Total P65/β-actin in glioma cell lines including (B-D) U-118 and (F–H) U-87. I&M Representative western blots of glioma cell lines treated with BSA or TNF for the indicated time points immunodetecting pSTAT3-Y705, total STAT3 and β-actin. β-actin was used as an internal control. Densitometry analysis of pSTAT3-Y705/β-actin, pSTAT3-Y705/Total STAT3 and Total STAT3/β-actin in glioma cell lines including (J-L) U-118 and (N-P) U-87. A two-way ANOVA Bonferroni’s multiple comparisons test was used when comparing 3 or more groups with 2 variables (* < 0.05, ** < 0.01, **** < 0.0001). h = hours