Fig. 5

PM zone enriched-Sec14l3 recruits PLC for activation upon Wnt5/Fz stimulation.

(A) Co-localization of Sec14l3 (green) with STIM1 proteins (red) in PC3 cells with or without Wnt5a stimulation. Arrows indicates PM-localized protein after Wnt5a stimulation. Scale bar, 10 μm. (B) Immunofluorescence of endogenous SEC14L2 in HEK293T cells with or without hFz5/mDvl2 transfection. Arrows indicates PM-localized SEC14L2 (green). Scale bar, 10 μm. (C) Immunofluorescence of Plcδ4a-mCherry (red), mDvl2-Flag (gray) and Sec14l3-GFP (green) in MCF7 cells. PM-localized Plcδ4a (red) are indicated by arrows. The bottom panel show the schematic representation of transfected constructs in corresponding rows and an interpretation of the results. Scale bar, 10 μm. (D) The Sec14l3 CARL-TRIO and GOLD domains are important for Rfz2 mediated Plcδ4a recruitment to the PM in the HEK293T cells. Statistical data from three independent experiments are presented as mean ± SEM on the right (*p<0.05; ns, non-significant; same for other statistical data below). (E) Rfz2-mediated Plcδ4a PM recruitment is abolished in stable SEC14L2-knockdown HEK293T cells and failed to be restored by Sec14l3-ΔNG overexpression. Statistical data are presented. (F) The Sec14l3 GOLD domain is important for Wnt5a mediated Plcδ4a recruitment to the PM in PC3 cells. Statistical data are presented. (G) SEC14L2 depletion perturbs Plcδ4a access to PIP₂. Equal amounts of purified Plcδ4a protein and liposomes with or without PIP₂ were incubated with control or SEC14L2 depleted cell lysates. Statistical data are presented. All numerical data represented as a graph in the figure are shown in Figure 5—source data 1.