Fig. 4

Knockdown of YAP expression suppresses cancer cell-induced PSC activation. Western blot (a) and Real-time PCR assays (b) were performed to evaluate the effect of YAP-shRNA on the expression of CTGF at the mRNA and protein levels in BxPC-3 and Panc-1 cells. c The representative IHC staining for YAP, CTGF and α-SMA in the NP and PDAC specimens, accompanied by strong YAP staining or with weak YAP staining and NP specimens from WT mouse and PDAC specimens from a KPC mouse model (bar, 100 μm). d A schematic diagram of indirect co-culture conditions. e A schematic diagram of direct co-culture conditions. f PSCs were treated with mixed conditioned medium from BxPC3-shNC or BxPC-3-shYAP cells for 48 h; then, the supernatant of PSCs was collected, and an ELISA was performed to evaluate the TGF-β1 level. g PSCs were treated with mixed conditioned medium from BxPC3-shNC or BxPC-3-shYAP cells for 48 h, and western blot assays were performed to evaluate the α-SMA and TGF-β1 levels. h PSCs were treated with mixed conditioned medium from BxPC3-shNC or BxPC-3-shYAP cells for 48 h, and cells were stained with an anti-α-SMA antibody (green) and counterstained with DAPI (blue) to identify nuclei. Representative images demonstrate that the α-SMA level of PSCs in conditioned medium from BxPC-3-shYAP cells was reduced significantly (bar, 50 μm). i PCs (BxPC-3-shYAP or BxPC3-shNC) and PSCs were cultured together for 48 h. Then, immunofluorescence analysis was performed to detect α-SMA (green) and CTGF (red) expression in the cells (bar, 100 μm). *P < 0.05