Fig. 4

CAF MSI2 deficiency decreases IL-6 secretion. A The presence of tumorigenic cytokines in the CM from Ctrl CAFs and gMSI2 CAFs was analyzed using a multiplex human cytokine array. Quantification of the cytokine profile was performed by densitometry using ImageJ software. Data are normalized to the positive control (n = 4; two biological replicates), averaged, and represented in a heatmap. See also Additional file 2: Fig. S9 for cytokine array dot blots. B Volcano plot of human cytokine antibody array in (A) shows differential expression of several soluble factors between Ctrl CAF-CM and gMSI2 CAF-CM. It is worth noting that IL-6 decreases the most upon depletion of MSI2 in CAFs (red dot). P < 0.05 are in yellow dots; two-tailed Student’s t-test. C IL-6 ELISA assessing and confirming lower IL-6 levels in gMSI2 CAF-CM versus Ctrl CAF-CM. Data are mean ± SD (n = 4); ^****P < 0.0001 versus Ctrl CAF-CM; two-tailed Student’s t-test. D Western blot analysis of IL-6 in gMSI2 CAFs in comparison to Ctrl CAFs. β-actin was used as a loading control. Data are mean ± SD (n = 3); ^***P < 0.001 versus Ctrl CAFs; two-tailed Student’s t-test. E Western blot analysis of IL-6 upon MSI2 overexpression with increasing amounts of plasmid (0 − 5 μg). Ctrl CAFs were transfected with pcDNA empty vector. F Correlation analysis of the protein levels of MSI2 and IL-6 in CAFs