Fig. 3

Coordinated expression and modulation of ZEB2, pCRAF and pASK1. a Left: immunoblot analysis of ZEB2, CRAF pS338, and ASK1 pS83 on whole lysates of SW480 cells treated for 4 days with 5 μM 5-fluorouracil (5-FU) or 2,5 μM oxaliplatin (OXA). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. Right: quantification of immunoblot shown on the left. b qRT-PCR analysis of ZEB2 levels in CCSCs (left panel) and SW480 (right panel) 24 h after siRNA-mediated silencing of ZEB2. ***P < 0.001 from two-tailed t test. Data of qRT-PCR are the mean ± SD, n = 3. c Immunoblot analysis of ZEB2, CRAF pS338, and ASK1 pS83 on whole cell lysates 24 h upon siRNA-mediated silencing of ZEB2 in CCSCs (left panel) and SW480 (right panel). The respective quantifications are shown on the right. d qRT-PCR analysis of ZEB2 levels in CCSCs (left panel) and SW480 (right panel) transduced with empty pLenti-GFP (Vector) or with pLenti-GFP-ZEB2 (ZEB2) and sorted on the basis of GFP expression. ***P < 0.001 from two-tailed t test. Data are the mean ± SD, n = 3. e Immunoblot analysis of ZEB2, CRAF pS338, and ASK1 pS83 on whole lysates of CCSCs (left panels) and SW480 cells (right panels) transduced with pLenti-GFP (Vector) or with pLenti-GFP-ZEB2 (ZEB2) and sorted as above. The respective quantifications are shown on the right. f Viability of CCSCs (left) and SW480 (right) transduced with pLenti-GFP or pLenti-GFP-ZEB2, sorted on the basis of GFP expression and immediately treated for 48 h with 10 μM oxaliplatin (OXA) and 10 μM 5-fluorouracil (5-FU). *P < 0.05 from two-tailed t test, n = 3. Data are the mean ± SD of three independent experiments