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. 2014 Dec 15;127(24):5303–5316. doi: 10.1242/jcs.157560

Fig. 5.

Fig. 5.

Eps8 is required for active Src localization to autophagosomes. (A) FAK WT and FAK−/− cells were transiently transfected with Eps8 siRNA (siEps8 #1 and #2; siControl, control siRNA), fixed and stained with anti-Eps8 and anti-Src pY416 antibodies. The quantification is representative of three independent experiments. Solid arrows indicate focal adhesions. Dashed arrows indicate active Src in autophagosomes. Scale bars: 20 µm. Results are mean±s.d. *P<0.01 (Student's t-test). (B) FAK−/− SCC cells were infected with either Eps8 (shE) or non-targeting (shC) shRNA and LC3B was immunoprecipitated (IP) from cell lysates. Samples were subjected to western blot analysis using anti-Eps8 and anti-Src pY416 antibodies. (C) FAK−/− cells were transiently transfected with two independent Eps8 siRNAs, fixed and stained with anti-Src pY416 antibody and TRITC–phalloidin. Solid arrows indicate active Src localization at focal adhesions. Dashed arrows indicate active Src in intracellular puncta. Scale bars: 20 µm. Insets show a magnified view of the boxed area.