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. 2018 Jun 29;29(8):2123–2138. doi: 10.1681/ASN.2017121312

Figure 5.

Figure 5.

GAPVD1 and ANKFY1 regulate podocyte migration rate. (A–E) Podocyte migration rate is analyzed by IncuCyte videomicroscopy. Representative images show human podocytes after induction of scratch wound (light blue) and 22 hours thereafter. Scratch wound area (light blue) and podocytes that have migrated (dark blue) are shown at 22 hours. Serum addition strongly increases podocyte migration rate. Podocytes stably expressing scrambled shRNA (negative control) show (A) complete wound closure, whereas (B–F) silencing GAPVD1 results in reduced migration. The decrease in podocyte migration was (C) strongly reversed by transfection of mouse Gapvd1 but (D and E) only partially rescued by murine Gapvd1 constructs reflecting mutations R937Q and L414V detected in patients with nephrotic syndrome. (F) Graph shows wound confluence versus time for conditions described in (A–E). Error bars indicate SD of 12 wells with identical conditions (n=3). (G–J) Podocyte migration is observed, indicating (G) complete wound closure upon stable expression of scrambled shRNA (negative control), whereas (H) silencing ANKFY1 in the presence of a mock-rescue results in reduced migration. The decrease in podocyte migration was (I) strongly reversed by transfection of shRNA-resistant ANKFY1, whereas (J) introduction of the mutation from family B1027 (R95L) into the rescue construct abrogates this ability. (K) Graph shows wound confluence versus time for the conditions described in (G–J). Error bars indicate SD of 12 wells with identical conditions (n=3).