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. 2022 May 9;221(7):e202109013. doi: 10.1083/jcb.202109013

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© 2022 Cail et al.

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Figure 5. — Curvature rescues a defect in endocytic site formation resulting from clathrin knockdown. (A and B) Representative images of control or clathrin siRNA cells grown on flat (A) or 75-nm ridge (B) substrate showing a marked increase in endocytic site fluorescence intensity and overlap of AP2 with dynamin2 in clathrin knockdown cells grown on nanoridge substrates. Scale bar, 10 μm. (C) Enrichment of AP2-RFP and DNM2-GFP is unchanged after clathrin knockdown, indicating that localization of CCPs to regions of induced membrane curvature is independent of clathrin expression. P values from multiple t tests, Mean ± SD, n = 3 cells per condition. (D) Induced curvature rescues the AP2 overlap with DNM2-GFP for automated AP2 detections; however, it does not rescue the AP2 overlap with residual CLTA-RFP detection for automated AP2 detections. ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, P > 0.05 from multiple t test. Mean ± SD, n = 3 cells per condition. (E) Model of endocytic protein stabilization in the absence of clathrin coats at regions of induced membrane curvature.