Figure 1

Verification of lnc-PCF as a novel lncRNA and its high expression level in the development of pulmonary fibrosis in vivo and in vitro. (a) In vitro translation assay showed that lnc-PCF performed no translation activities. The black arrow indicates that the positive control could translate a 75 kDa protein. (b) RNA FISH detecting the location of endogenous lnc-PCF (red) in cells. The result showed that lnc-PCF mainly localized in the cytoplasm. U6 and 18S RNA was used as nuclear and cytoplasmic localization markers, respectively. DNA (blue) was stained with DAPI. (c) Nucleocytoplasmic separation result confirmed that lnc-PCF was mainly expressed in the cytoplasm by using qRT-PCR. (d) Increased lnc-PCF expression in BLM-treated lung tissues at 0–28 days by using qRT-PCR. (e) Hydroxyproline (Hyp) content increased in the rat pulmonary tissues at 0–28 days, and it was higher at 21–28 day BLM-treated groups than that in the normal group. (f) Positive correlation was measured between the lnc-PCF expression and Hyp content in vivo by using Pearson correlation coefficient. (g) Increased lnc-PCF expression in TGF-β1-induced rat lung epithelial-t-antigen negative (RLE-6TN) cells from 0 h to 72 h. (h) Cells were transfected with lnc-PCF smart silencer /NC. And The smart silencer could significantly inhibit the expression of lnc-PCF. NC indicates a negative control of lnc-PCF smart silencer. (i) Recombinant plasmid of overexpressed lnc-PCF could increase the expression level of lnc-PCF. BP indicates blank plasmid. RP indicates recombinant plasmid of the overexpressed lnc-PCF. Each bar represents the mean±S.D., n=6. *P<0.05, **P<0.01