Fig. 5

Downregulation of OGT effectively induced autophagy, which had a pro-survival role in human bladder cancer cells. (a) Expression of autophagy markers (LC3, P62, and Beclin 1) was examined using the Western blot analysis. T24 (b) and UMUC3 (c) cells were transfected with si-OGT, and with or without 10 μM CQ, for 24 h, and the protein levels of LC3 were assessed using the Western blot analysis. Transfected T24 and UMUC3 cells were transfected with the GFP-LC3 construct and the transfectants were treated with or without CQ (10 μM) for 24 h. (d) Representative images of GFP-LC3 puncta were captured using a confocal fluorescence microscope (magnification, × 400). (e and f) Number of puncta per GFP-LC3-positive cell was calculated and presented. (g and h) T24 cells were transfected with si-OGT or si-NC, and with or without 10 μM CQ for 24 h. Apoptosis was measured using flow cytometry, and cell viability was examined using MTT assay (I)