Fig. 4

circPARD3 inhibits autophagy of LSCC cells. a FD-LSC-1 and Tu 177 cells were infected with circPARD3 overexpression lentiviruses (circPARD3-OE) or control empty vector (Vector). Expression of circPARD3 was examined by qPCR. b FD-LSC-1 and Tu 177 cells were transfected with siRNAs targeting circPARD3 (si-circ-1 and si-circ-2) or NC siRNAs (si-NC) for 48 h. Expression of circPARD3 was examined by qPCR. c and d FD-LSC-1 and Tu 177 cells were infected with circPARD3 overexpression lentiviruses or transfected with siRNAs targeting circPARD3. Western blotting was performed to detect the expression of LC3B and p62 after overexpression (c) or silencing of circPARD3 (d). e and f mCherry-EGFP-LC3B labeled FD-LSC-1 and Tu 177 cells were infected with circPARD3 overexpression lentiviruses or transfected with siRNAs targeting circPARD3. Autophagic flux was analyzed by confocal microscopy after overexpression (e) or silencing (f) of circPARD3. Scale bar, 25 μm. g qPCR analysis of circPARD3 in FD-LSC-1 and Tu 177 cells after Rapamycin (100 nM for 24 h) or 3-Methyladenine (3-MA, 2.5 mM for 24 h) treatment. Error bars represent SD of three independent experiments. *P < 0.05, **P < 0.01. N.S., no significant