Fig. 2

ALKBH5 regulated the autophagy flux. a The expression of ALKBH5 mRNA in ovarian cancer cells was determined using qPCR. b The expression of LC3-II and ALKBH5 was detected using WB. c SKOV3 cells infected with LVRU6P-01, LVRU6P-02, or LVRU6P-NC. After 48 h, puromycin was added at a concentration of 2.5 mg/mL. Then, cells were transfected with a plasmid encoding mRFP-GFP-LC3 that was distributed in SKOV3 cells and analyzed by confocal microscopy after 48 h post-transfection. The LC3 puncta were quantified using Image Pro-Plus 6.0 software. All experiments were repeated three times. The bottom panel indicates the quantification of LC3 punctate staining. d A2780 cells were infected with LV121-NC or LV121-ALKBH5. After 48 h, puromycin was added at a concentration of 2.5 mg/mL. Then, cells were transfected with a plasmid encoding mRFP-GFP-LC3. The distribution of mRFP-GFP-LC3 in A2780 cells was analyzed by confocal microscopy after 48 h post-transfection. The LC3 puncta were quantified using Image Pro-Plus 6.0 software. All experiments were repeated three times. The bottom panel indicates the quantification of LC3 punctate staining. e and f The expression of LC3-II was detected by WB. Error bars represent the standard error. * indicated P < 0.05. Scale bar: 5 μm