Fig 2: Overexpression of miR-31-5p directly targeted LIN28B to repress the development of HG-stimulated HMCs. A The protein level of LIN28B in HMCs transfected with pcDNA or LIN28B was measured via western blot assay. B–I HMCs were transfected with miR-NC, miR-31-5p, miR-31-5p + pcDNA or miR-31-5p + LIN28B in HG condition. B The protein level of LIN28B in HMCs was measured using western blot assay. C HMC viability was assessed by CCK-8 assay. D The concentrations of IL-6 and TNF-α in HMCs were examined with ELISA kits. E Cell cycle in HMCs was analyzed by flow cytometry analysis. F HMC proliferation was evaluated by EdU assay. G–I The protein levels of CyclinD1, P21, collagen I, collagen IV and TGF-β1 in HMCs were measured by western blot assay. J and K The activity of SOD and the level of MDA in HMCs were examined with commercial kits. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Fig 3: Lin28B was upregulated in colon cancer tissues, as detected by immunohistochemical staining (magnification, ×100). The magnification in the top-right corner of the images is ×400. (A) Lin28B was not expressed in adjacent colon tissue. (B) Lin28B was expressed in colon cancer tissue. Lin28B, Lin-28 homolog B.

Fig 4: Overexpression of Lin28B promotes the chemotherapeutic sensitivity of colon cancer cells to 5-FU. (A) Lin28B was predominantly located in the cytoplasm of HCT116 cells, as illustrated by GFP distribution, and the nucleus was stained by DAPI (magnification, ×200). (B) Overexpression of Lin28B was confirmed by quantitative polymerase chain reaction. (C) Overexpression of Lin28B was confirmed by western blot analysis. (D) The overexpression of Lin28B significantly increased the chemosensitivity of HCT116 cells treated with 5-FU (P<0.05). Lin28B, Lin-28 homolog B; 5-FU, 5-fluorouracil; GFP, green fluorescent protein.

Fig 5: MiR-31-5p directly targeted LIN28B. A LIN28B contained miR-31-5p binding sites. B and C Dual-luciferase reporter assay and RIP assay were conducted to analyze the interaction between miR-31-5p and LIN28B. D The mRNA level of LIN28B in the serums of DM, DN patients and healthy volunteers was detected by RT-qPCR assay. E The correlation between the levels of mIR-31-5p and LIN28B in DN patients’ serums was analyzed by Spearman’s correlation coefficient analysis. F The protein level of LIN28B in HG-treated HMCs was measured via western blot assay. G The protein level in HG-treated HMCs transfected with miR-NC, miR-31-5p, anti-miR-NC or anti-miR-31-5p was measured through western blot assay. ***P < 0.001, ****P < 0.0001