Figure 4.

The expression of ATG14 affected cell apoptosis and autophagy. (A) TCGA Nature 2014 and TCGA Provisional database analyses showing the correlation between EIF3J-DT and ATG14 mRNA expression levels. (B) mRNA expression level of ATG14 in the MGC803 cells with overexpressed (left), silenced (middle), or overexpressed+silenced (right) EIF3J-DT detected by qPCR. (C) Cell viability among the NC, siATG14 #1, and siATG14 #2 MGC803 cells 24 h after being treated with OXA (left) and 5-Fu (right) detected by MTT assay. (D) Cell viability among the NC, siATG14 #1, and siATG14 #2 drug-resistant MGC803 cells 24 h after being treated with OXA (left) and 5-Fu (right) detected by MTT assay. (E) Numbers of colony formation of the NC, siATG14 #1, and siATG14 #2 MGC803 (left) and MKN45 (right) cells 14 d after being treated with PBS, OXA (10 μg/mL) and 5-Fu (20 μg/mL). (F) Apoptosis rates of the NC, siATG14 #1, and siATG14 #2 MGC803 (left) and MKN45 (right) cells 24 h after being treated with PBS, OXA (10 μg/mL) and 5-Fu (20 μg/mL) detected by flow cytometry. (G) Western blotting performed on the NC, siATG14 #1, and siATG14 #2 MGC803 cells 24 h after being treated with OXA (20 μg/mL) (left) and 5-Fu (20 μg/mL) (right) against C-PARP, PARP, C-CASP3, CASP3, C- CASP7, and CASP7. (H) Expression levels of ATG14, LC3-I, LC3-II and SQSTM1 in the NC, siATG14 #1, and siATG14 #2 MGC803 (left) and MKN45 (right) cells detected by western blotting. (I) Expression levels of LC3-I and LC3-II in the NC, siATG14 #1, and siATG14 #2 MGC803 cells with or without CQ treatment (10 μM) detected by western blotting. (J) Representative fluorescence images of the GFP-mCherry-LC3 transfected NC and siATG14 (siRNA2) MGC803 cells 24 h after being treated with PBS, OXA (10 μg/mL) and 5-Fu (20 μg/mL) (left). Scale bar: 10 μm. The statistics was shown on the right. Data were quantified as mean ± SEM (n = 3). **P < 0.01, ***P < 0.001, vs the relative control