Research Paper Volume 16, Issue 2 pp 1145—1160

Figure 4. The effect of BECN2 knockdown on the function of ATDC5. (A) The effect of siRNA-BECN2 on the mRNA expression of BECN2 was detected by RT-PCR. (B) The protein level of BECN2 was evaluated by western blot. (C) The protein expression of BECN2 were determined using ImageJ software, GAPDH was used as the internal control, respectively (n=3). (D) CCK-8 was used to assess cell viability. (E, F) Cell apoptosis was detected by Muse. (G) The immunofluorescence staining of LC3 was used to evaluate the autophagy level (green signals represent LC3, blue signals represent DAPI, scale bar: 10 μm). (H, I) Autophagy-related protein expression levels were determined by immunoblotting. (J, K) The expression of IL-1β, Caspase1 and IL-18 were assessed by western blot. Relative protein expression was qualified by ImageJ software, GAPDH was used as the internal control, respectively. All data represent mean ± SD. All in vitro experiments were repeated three times independently. * p < 0.05, ** p < 0.01, and *** p < 0.001.