Fig. 8

lnc-Helf promotes hepatic inflammation and fibrosis by interacting with PTBP1 to facilitate PIK3R5 mRNA stabilization. A The RNA level of PIK3R5 was detected in liver samples of fibrotic patients (n = 28) and healthy people (n = 6) by qRT–PCR analysis. B The correlation of lnc-HELF and PIK3R5 was assessed by Pearson correlation analysis, n = 34. C, D RIP–qRT–PCR assay in single-cell suspensions from the liver (C) and RAW264.7 cells transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 (D). E The expression and location of PTBP1 in primary HSCs infected with lenti-control or lenti-lnc-Helf was assessed by confocal microscopy. Scale bar is 20 μm. The fluorescence intensity of nuclear and cytoplasm was quantified on the right. F qRT–PCR was used to detect the RNA level of Pik3r5 in lnc-Helf-overexpressed primary HSCs simultaneously transfected with siPTBP1. G RAW264.7 cells transfected with pcDNA3.1 or pcDNA3.1-lnc-Helf with or without si-PTBP1 were treated with 5 μg/ml actinomycin D. RNA of each group was extracted at indicated time points (0, 2, 4 h) and the expression of Pik3r5 mRNA was analyzed by qRT–PCR and normalized to Gapdh. Data are presented as mean ± SEM. ^*/#p < 0.05. *p < 0.05 for IgG RIP or LV-control + si-NC or pcDNA3.1 + si-NC. ^#p < 0.05 for LV-lnc-Helf + si-NC or pcDNA3.1-lnc-Helf + si-NC, unpaired Student’s t test (A, C, E) and one-way ANOVA (D, F, G)