Fig 1: Loss of VPS34 resulted in abnormal assembly of retromer complexes in oocytes. MII oocytes were collected from 5 W control and ZcKO mice after superovulation. a‐e) Immunofluorescent co‐staining of VPS26A (red) with LAMP1 (green) (a); RAB7 (red) with VPS26A (green) (b); RAB7 (red) with VPS35 (green) (c); RAB7 (red) with TBC1D5 (green) (d), M6PR (red) with LAMP1 (green) (e) in control and ZcKO oocytes. Scale bars = 50 µm. Blue, nuclear staining with Hoechst 33342; Merge, representing a superposition of fluorescence signals from three channels. f) Western blot of VPS34 and CTSD and M6PR in control and ZcKO oocytes. β‐tubulin was used as an internal control. g) Western blots of TBC1D5, VPS35, VPS26A and VPS26B in control and ZcKO oocytes of mice. β‐tubulin was used as a loading control. h) Immunofluorescent co‐staining of VPS26B (red) with LAMP1 (green) in control and ZcKO oocytes. i–m) Interactions of RAB7 with retromer components in control and Vps34 knockout (KO) 293T cells. Vps34 knockout 293T cell line was constructed using the CRISPR‐Cas9 system and co‐immunoprecipitation experiments were performed as follows: RAB7 (i), TBC1D5 (j), VPS26B (k), VPS26A (l) and VPS35 (m) were used to immunoprecipitate the proteins in control and KO 293T cells, and then anti‐RAB7, anti‐TBC1D5, anti‐VPS26A, anti‐VPS26B and anti‐VPS35 antibodies were used for Western blot detection. At least 3 independent replicates were performed for each experiment. n) Direct interaction between VPS34 and RAB7 by GST pull‐down assay.