Figure 2. Effects of hsa_circ_0070963 overexpression on HSCs. LX2 cells were transduced with hsa_circ_0070963 for 48 h. (A) hsa_circ_0070963 mRNA levels in transduced LX2 cells. (B, C) HSC proliferation was detected using the 5-Ethynyl-20-deoxyuridine (EdU) assay (B) and the IOD values are shown (C). (D, E) Northern blot analysis of hsa_circ_0070963 expression after RNase R treatment. (F–H) Cell cycle analysis was performed with control or hsa_circ_0070963-overexpressing cells and the cell cycle distribution is shown. (I, J) The mRNA levels of α-SMA (G) and Col1A1 (H) were measured by qRT-PCR. (K) The protein levels of α-SMA and Col1A1 were measured by western blot. (L) Immunofluorescence staining for α-SMA (green) and Col1A1 (green) was evaluated by confocal laser microscopy. DAPI was used to stain the nuclei. The scale bar represents 50 μm. Data are presented as means ± SD of three experiments (*p < 0.05 and **p < 0.01).