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. 2018 Nov 5;217(11):3817–3828. doi: 10.1083/jcb.201804132

Figure 4.

Figure 4.

Defective autophagosome formation in TMEM41B-KO and VMP1-KO cells. (A) WT, TMEM41B-KO, VMP1-KO, and rescued TMEM41B-KO HEK293T cells were cultured under nutrient-rich and starvation (Stv) conditions for 2 h and subjected to immunofluorescence microscopy. Bars: 10 µm (main images); 500 nm (insets). (B) Numbers of FIP200 and WIPI2 puncta in cells under nutrient-rich and starvation conditions were quantified as in Fig. 3 E. Data were collected from >70 cells for each sample. (C) Quantification of colocalization between FIP200 and LC3 and between WIPI2 and LC3. Data were collected from >70 cells for each sample. (D) WT, TMEM41B-KO, VMP1-KO, and rescued TMEM41B-KO cells expressing mRuby3-STX17TM were starved for 2 h and observed by fluorescence microscopy. Data were collected from >40 cells and quantified as in Fig. 3 E. Bars: 10 µm (main images); 500 nm (insets). (E) Transmission EM of WT, VMP1-KO, and TMEM41B-KO cells under starvation conditions (2 h). Insets indicate an autophagosome (in WT) or ferritin clusters (in KO). Bars: 500 nm (main images); 50 nm (insets). (F) WT, TMEM41B-KO, VMP1-KO, and rescued TMEM41B-KO HEK293T cells were stained with LipidTOX. Bar, 10 µm.