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. 2018 May 29;38(12):e00102-18. doi: 10.1128/MCB.00102-18

FIG 5.

FIG 5

PTPL1 binding is required for PTEN-dependent apical membrane clustering. (A) Schematic representation of PTPL1 mutants. KIND, kinase noncatalytic C-lobe domain; FERM, 4.1, ezrin, radixin, moesin domain; PTP, protein tyrosine phosphatase domain. (B) Coimmunoprecipitation of V5-PTEN with GFP-PTPL1 wt and PDZ deletion mutants in HEK293T cells. Bottom, quantification of relative V5-PTEN binding to GFP-PTPL1. Error bars represent the SEM (n = 4). (C) Quantification of apical membrane size in W4:shCTRL and W4:shPTPL1 cells expressing EV, GFP-PTPL1 wt, or PDZ deletion mutants. Red bars represent the average. Error bars represent the SEM (n > 30 in at least three experiments). *, P < 0.05 using independent sample t tests. ns, not significant (P > 0.05). EV, empty vector. (D) Localization of GFP-PTPL1, GFP-PTPL1(ΔPDZ1-5), and Lifeact-Ruby in PTPL1-depleted W4 cells. Scale bars, 5 μm. (E) Coimmunoprecipitation of Flag-PKN2 with GFP-PTPL1 wt and PDZ deletion mutants in HEK293T cells. (F) Coimmunoprecipitation of Flag-ArhGAP29 with GFP-PTPL1 wt and PDZ deletion mutants in HEK293T cells.