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. 2020 Nov 20;8:515051. doi: 10.3389/fcell.2020.515051

FIGURE 4.

FIGURE 4

Upregulation of autophagy promoted the invasion and EMT progression of si-NDUFA4L2-transfected 143b and HOS cells. 143b and HOS cells were treated with Rapamycin post-transfection with si-NC or si-NDUFA4L2 in hypoxic environments. (A) Western blotting was performed to determine protein expression of P62, LC3, E-cadherin, and Vimentin in 143b cells. For P62, si-NC vs. si-NC + Rapamycin was #p < 0.001, si-NC vs. si-NDUFA4L2 was #p < 0.001, si-NDUFA4L2 vs. si-NDUFA4L2 + Rapamycin was ∗p < 0.05; For LC3, si-NC vs. si-NC + Rapamycin was #p < 0.001, si-NC vs. si-NDUFA4L2 was #p < 0.001, si-NDUFA4L2 vs. si-NDUFA4L2 + Rapamycin was #p < 0.001; For E-cadherin and Vimentin, si-NC vs. si-NC + Rapamycin was nsp ≥ 0.05, si-NC vs. si-NDUFA4L2 was # < 0.001, si-NDUFA4L2 vs. si-NDUFA4L2 + Rapamycin was # < 0.001. (B) Immunofluorescence was performed to measure E-cadherin and Vimentin expression in 143b cells. (C) Transwell assays were performed to determine the invasion ability of si-NDUFA4L2-transfected 143b and HOS cells post-treatment with Rapamycin. HY: hypoxic environment. nsp ≥ 0.05, ψ < 0.01, and # < 0.001 were defined as measures that indicated significant differences among treatment groups. All experiments were performed in triplicate.