Fig. 3: PR55α-knockdown by shRNA inhibits YAP and activates the MOB1-LATS1/2 cascade in pancreatic cancer cells. | Oncogenesis

Fig. 3: PR55α-knockdown by shRNA inhibits YAP and activates the MOB1-LATS1/2 cascade in pancreatic cancer cells.

From: PR55α regulatory subunit of PP2A inhibits the MOB1/LATS cascade and activates YAP in pancreatic cancer cells

Fig. 3

a CD18/HPAF and AsPC-1 cells were stably transduced with Dox-inducible shRNAs targeting various regions of PR55α or, as a control, with nontargeting shRNA. After incubation with Dox (2 µg/ml) for 3 days, the cell lysates (100 µg) were analyzed by immunoblotting for the indicated protein levels and/or phosphorylation. GAPDH served as an internal protein expression control. b Validation of the effects of PR55α on MOB1/LATS/YAP cascade in pancreatic cancer cells. shRNA-transduced CD18/HPAF cells were incubated with Dox (2 µg/ml) for the days indicated and analyzed for PR55α expression and the levels/phosphorylation of MOB1, LATS1/2, and YAP. GAPDH serves as an internal control. c To test the effect of PR55α on LATS2 protein stability, shRNA-transduced CD18/HPAF cells were incubated in medium containing cycloheximide (CHX, 15 μg/ml) to halt protein synthesis for the indicated hours. Whole-cell extracts were analyzed for LATS2 and GAPDH protein levels by immunoblotting. The protein levels of LATS2 and GAPDH were quantified using ImageJ software; relative LATS2 levels were normalized with GAPDH levels in the samples and protein half-life determined using SigmaPlot (version 11.2) analytical program

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