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Fig. 3 | BMC Cell Biology

Fig. 3

From: ULK1 phosphorylates Sec23A and mediates autophagy-induced inhibition of ER-to-Golgi traffic

Fig. 3

Autophagy and ULK1 caused retention of transport marker VSVG in ERESs. a Hela cells were transfected with GFP-VSVG-tsO45 and the cells were kept at 39.5C overnight. Then the cells were cultured in (a) normal growth medium or (b) induced with autophagy by amino acid starvation for 2 h before the incubation temperature was shifted to 32 °C for the indicated time to chase the VSVG out of the ER. c Quantification of the ERES-associated VSVG signals. The fractions of VSVG signal in ERES at different time points after 32 °C chase were measured by Manders Coefficients. Number of cells ≥10; Error Bars = S.D. d Hela cells were transfected with GFP-VSV-G tsO45 and Myc-ULK1 or (e) its kinase dead mutant at 1:4 ratio and the cells were kept at 39.5 °C. After being chased at 32 °C for different times as indicated, the cells were fixed and stained with Sec31A for ERES. f Quantification of the amount of ERES-retained VSVG signal in ULK1 or kinase dead mutant. The fractions of VSVG signal in ERES at different time points after 32 °C chase were measured by Manders Coefficients. Number of cells ≥ 8; Error Bars = S.D., p < 0.05

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