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. 2016 Jul 8;6:29256. doi: 10.1038/srep29256

Figure 5. Autophagy is involved in the clearance of accumulated EGFR.

Figure 5

(a) The cells were pretreated with 10 mM 3-MA or 20 μM 6r for 1 h and then stimulated with 100 ng/mL EGF for additional periods of 2 and 6 h. Subsequently, the cells were harvested and subjected to SDS-PAGE and Western blotting. EGFR degradation was examined through immunostaining by using an anti-EGFR antibody. ACTIN signalling was considered the loading control. (b) After 24 h of shRNA knockdown of CTSS, the cells were treated with 100 ng/mL EGF for the indicated durations. The harvested cells were subjected to Western blotting. (c) The cells were pretreated with 20 μM 6r for 1 h and then treated with 100 ng/mL EGF for 30 min. The cells were immediately fixed, permeabilised, stained with EGFR and LC3 antibodies, as described in Methods.