Fig. 6. NRP2 isoforms differentially affect MET activation.

(A) Western blotting for MET-signaling markers in response to HGF (h, hours) in serum-starved H358 cells bearing the indicated NRP2 knockdown, without or with reexpressed isoforms. (B) Left: TGFβ-induced migration of H358 cells bearing NRP2 knockdown and reexpressed isoforms was assessed in the presence or absence of the MET inhibitor, PHA-665752 (PHA). Data were analyzed as in Fig. 2, A and C. Dox induction was omitted in these experiments, accounting for the reduced NRP2b-mediated migration rescue. n = 3 experiments; linear combinations of ANOVA parameters analyzed on a log scale. (f) P > 0.05 for these pairwise comparisons in the ANOVA model. However, in stand-alone t tests, P = 0.0006 and 0.0003, respectively. The results for shCtl and shNRP2 with reex-pressed NRP2b had lower variability compared to shNRP2 alone and shNRP2 with reexpressed NRP2a. Thus, their comparisons were adversely affected by pooling errors in the ANOVA model, which by default assumes equal variance among groups. Right: Western blot confirmation of MET inhibition. Blots are representative of three independent experiments.