Fig 2: Doxazosin inhibits autophagy by activating the PI3K/Akt/mTOR signaling pathway in LX-2 cells. The protein levels of p-PI3K, total PI3K, p-Akt, total Akt, p-mTOR, and total mTOR in LX-2 cells treated with the indicated concentrations of doxazosin (A) for the indicated times (B) were measured by immunoblotting. LX-2 cells were treated with the indicated chemicals. The protein levels of p-mTOR, total mTOR, Bax, Bcl-2, p62, LC3B, COL1A1 and α-SMA were revealed by immunoblotting (C), while those of α-SMA, LC3B and p62 were revealed by immunofluorescence ((D), Scale bar, 50 μm), and apoptosis was evaluated by flow cytometry (E).

Fig 3: Expression of mTOR and HIF-1a protein. a-b Western blotting was used for the analysis. c Expression of proteins in the mouse gastric. Data are expressed as the mean ± SD. ** indicates P < 0.01 compared with the control group. ## indicates P < 0.01 compared with the model group. n = 5 in each group

Fig 4: MiRNA-138-5p targets the PTK2 expression involved in AKT/mTOR signal pathway.A Bioinformatic analysis of the potential target gene by prediction software(PicTar, miRTargetbase, Targetscan, miRDB). B TACG data base analysis of the expression level of PTK2 in HCC. C The person analysis the miRNA-138-5p and PTK2 expression correlation in HCC tissues. D, E The q-RT-PCR and western-blot assay analysis the PTK2 expression in miRNA-138-5p inhibition and overexpressed HCC cells. F The bioinformation analysis the binding site of miRNA-138-5p and PTK2. G, H The luciferase activity assay was used to verify the combination of miRNA-138-5p and PTK2. I The western blot analysis miRNA-138-5p targets the PTK2 expression involved in PI3K/AKT signal pathway. Data are shown as the mean ± standard deviation of three independent experiments. *P < 0.05was regarded as statistically significant.

Fig 5: Knockdown of circC16orf62 inhibited in vivo growth of HCC cell.A The BALB/c nude mice were sacrificed for the xenografts, and the size was measured by the beside ruler. B The tumor growth curve of xenografts was plotted in shNC and shcircC16orf62 group (n = 5 each group) by measuring the tumor size (width2 × length × 0.5) with vernier caliper. C The anatomized subcutaneous tumor xenografts were weighed and analyzed with student’s t test. D IHC analysis the proliferation markers (PCNA and KI-67) in different group xenografts, the scale bar represents 100 µm. E IHC analysis the AKT/mTOR signaling pathway markers in different group xenografts, the scale bar represents 100 µm. Data are shown as the mean ± standard deviation of three independent experiments. *P < 0.05 was regarded as statistically significant.