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. Author manuscript; available in PMC: 2014 Nov 1.
Published in final edited form as: Nat Cell Biol. 2014 Apr 6;16(5):401–414. doi: 10.1038/ncb2934

Figure 7. Cx43 modulates formation of pre-autophagosomal structures through direct interaction with autophagy-related proteins.

Figure 7

(a) Co-immunostaining for Cx43 and the indicated Atgs in NRK cells maintained in the presence or absence of serum (top). Single and merged channels of the 3D reconstruction of the boxed regions are shown (bottom). Nuclei are highlighted with DAPI. (b, c) Immunoblot for the indicated proteins of immunoprecipitates (IP) of Cx43 in NRK cells maintained in the presence or absence of serum (b) or treated or not with 3-methyladenine (c) Atg5 is shown as negative control for IP. (d) Co-staining for Atg9 and e-cadherin in NRK cells control and knocked down for Cx43. Single and merged channels at higher magnification areas are shown. (e) Immunofluorescence for endogenous Atg9 and Cx43 in NRK cells control or knocked down for Eps15. (f, g) Immunofluorescence for endogenous Cx43 (green) in NRK cells control or knocked down for Atg14 (f) or Atg9 (g) maintained in the presence or absence of serum. (h) Immunofluorescence of endogenous Cx43 in NRK cells control or knocked down for Atg14 treated (or not) with Tx or Lnd (left). e-cadherin staining (red) is shown to highlight plasma membrane in g and h. Full fields are shown in Supplementary Fig. 6h. Quantification of intracytoplasmic Cx43-positives vesicles (right) (n= 3 wells, 3 independent experiments, >30 cells per experiment). Values are mean+s.e.m and significant for (*) p<0.01 using ANOVA+Bonferroni test. Bars: 5 μm. Uncropped images of blots are shown in Supplementary Fig. 9.