Figure 3

lnc-PCF promoted the proliferation of epithelial cells activated with TGF-β1. (a) lnc-PCF smart silencer evidently inhibited cell proliferation compared with the TGF-β1+NC group. Proliferation analysis was performed using an xCELLigence real-time cell analysis (RTCA) system. (b) PCNA protein levels were analyzed by Western blot analysis. The lnc-PCF smart silencer evidently retarded PCNA expression compared with that in the TGF-β1+NC group. (c) Knockin lnc-PCF could promote cell proliferation compared with the normal control group. Proliferation analysis was performed using a RTCA system. (d) Knockin lnc-PCF could promote the proliferating cell nuclear antigen (PCNA) expression compared with that of the normal control group by western blot analysis. (e) Cell cycle was analyzed by flow cytometry. Cells were pretreated with 50 nM lnc-PCF smart silencer/NC for 6 h and then exposed to 5 ng/ml TGF-β1 for 72 h. The DNA in RLE-6TN cells were labeled with propidium iodide (PI). (f) Percentage histogram of cell cycle distribution was presented according to flow cytometry analysis. Lnc-PCF smart silencer increased the percentage of cells in the G1/G0 phase and decreased that in S and G2/M. (g) Cell cycle was analyzed by flow cytometry. Cells were pretreated with 2 μg of BP/RP for 6 h and subsequently exposed to 5 ng/ml TGF-β1 for 72 h. The DNA in RLE-6TN cells was labeled with PI. (h) Percentage histogram of cell cycle distribution was presented according to flow cytometry analysis. Knockin lnc-PCF reduced the percentage of cells in the G1/G0 phase and increased that in S and G2/M. (i) Lnc-PCF smart silencer decreased and increased the expression levels of cyclin E and cyclin B, respectively, as shown by western blot analysis. (j) Knockin lnc-PCF increased and inhibited the expression levels of cyclin E andcyclin B, respectively, as shown by western blot analysis. NC indicates a negative control of lnc-PCF smart silencer, BP indicates blank plasmid, and RP indicates the recombinant plasmid of overexpressed lnc-PCF. Each bar represents the mean±S.D., n=6, *P<0.01, **P<0.05