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. 2013 Dec 16;124(1):338–352. doi: 10.1172/JCI71526

Figure 7. TGF-β is a driver of Wnt7b expression in KRC cells.

Figure 7

(A) KRC cells (white bars) expressed high Wnt7b mRNA levels compared with KC (control, black bars; *P < 0.04) and KSC (gray bars; *P < 0.02) cells. (B and C) TGF-β1 (0.5 nM) increased Wnt7b mRNA (B, white bars; *P = 0.032) and protein (C) levels in KRC cells. (C) Representative blot from two independent experiments. ERK2 was used to confirm equivalent lane loading. (D and E) SB505124 (D, 2 μM, white bars) and anti–TGF-β2 (E, 5 mg/ml, white bars) reduced Wnt7b mRNA levels in KRC cells (**P < 0.001) compared with their respective controls (black bars). (F) TGF-β1 (0.5 nM, white bars) did not alter Wnt7b mRNA levels in KSC cells. (G) TGF-β1 (0.5 nM) increased wild-type Wnt7b promoter activity (left panel) in KRC cells (*P = 0.002), whereas KSC cells had low basal activity that was not increased by TGF-β1. TGF-β1 failed to increase the activity of an SBE-mutated Wnt7b reporter in KRC cells (right panel). (H) ChIP assays revealed that Smad2-4 (Smad) bound the Wnt7b promoter in TGF-β1–stimulated (0.5 nM) KRC cells. Left panel: quantification; **P < 0.001. Right panel: representative gel image from three independent experiments. (I) hPDAC PCCs with strong cytoplasmic Wnt7b immunoreactivity showed nuclear Smad4 (top panels), whereas hPDACs that lack Wnt7b did not demonstrate Smad4 immunoreactivity (bottom panels). (I) Representative images from two PDACs. Scale bars: 50 μm. (A, B, and D–H) Data represent the means ± SEM from three independent experiments.