Journal: Biochemistry and Biophysics Reports

Article Title: LncRNA RMST knockout inhibits fibrosis by down-regulating Smad3 during mouse skin wound healing

doi: 10.1016/j.bbrep.2025.102386

Figure Lengend Snippet: RMST-KO reduces fibrosis in mouse skin wound healing. (A) Differences of RMST expression between unwounded skin and the 21 dps group. (B) Agarose gel electrophoresis confirmed the successful knockout of RMST. (C) QPCR assay showing the expression of RMST 21 dps. (D) H&E and Masson staining of the wounds 21 dps as indicated. (E–F) RMST-KO reduced the (E) thickness and (F) area of neogenic fibrous tissue at 21 dps. (G–H) QPCR showing the COL1A1 and α-SMA levels were significantly decreased in the RMST-KO group compared to the WT group 21 dps. (I) Representative blot of COL1A1 and α-SMA 21 dps. (J–K) Statistical analysis of the immunoblot bands indicated that RMST-KO significantly suppressed the expression of COL1A1 and α-SMA 21 dps. Data were shown as mean ± SD, n = 6, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: α-SMA (67735-1-Ig) , 1:50000 , Proteintech.

Techniques: Expressing, Agarose Gel Electrophoresis, Knock-Out, Staining, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: LncRNA RMST knockout inhibits fibrosis by down-regulating Smad3 during mouse skin wound healing

doi: 10.1016/j.bbrep.2025.102386

Figure Lengend Snippet: Smad3-OE reversed the fibrosis suppression resulting from RMST-KO. (A) QPCR confirmed the upregulating effect of Smad3-OE lentivirus. (B) Representative blot of Smad3 under the effect of Smad3-OE and/or RMST-KO. (C) Statistical analysis of the immunoblot bands. (D) H&E and Masson staining of the wounds as indicated. (E–F) Statistical analysis of the (E) thickness and (F) area of neogenic fibrous tissue 21 dps. (G–I) qPCR detection of fibrosis markers and proteins in classical fibrosis pathways, including COL1, α-SMA, and TGF-β1. (J–M). Representative blot of COL1, α-SMA, and TGF-β1. (N–O) qPCR detection of inflammatory cytokines TNF-α and IL-1β. (P–R) Representative blot of TNF-α and IL-1β. Data were shown as mean ± SD, n = 6, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: α-SMA (67735-1-Ig) , 1:50000 , Proteintech.

Techniques: Western Blot, Staining

Journal: Cell Death & Disease

Article Title: Targeting sorting nexin 3 to treat pulmonary fibrosis by dual modulating Wnt/β-catenin signaling

doi: 10.1038/s41419-025-08248-x

Figure Lengend Snippet: A The protein mass spectrum of Wls were shown. B Fresh lung homogenates were sparked by anti-Flag or anti-Wls for Wls or SNX3 detection in co-IP analysis. C IHC staining analysis shown the Wls expression in BLM-induced mice (Scale bar: 200 μm; n = 8 mice). D The protein level and colocalization of Wls with SNX3 in Snx3-cTg mice were detected by IF staining analysis (Scale bar: 100 μm, n = 8 mice). E The protein level of Wls were detected by IHC staining analysis in Snx3-cTg mice (Scale bar: 200 μm; n = 8 mice). F The protein level and colocalization of Wls with SNX3 in Snx3-cKO mice were detected by IF staining analysis (Scale bar: 100 μm, n = 8 mice). G The protein level of Wls in Snx3-cKO mice were detected by western blotting analysis ( n = 8 mice). H , I IF staining analysis were performed to detected the protein level and colocalization of Wls with Rab 5a and LAMP1; Scale bar: 10 μm; n = 3 experiments. J The Wls intracellular recycling and degradation flow chart. Were created with BioGDP.com K H&E staining and PSR staining in lung tissue sections were shown. Scale bar: 200 μm, n = 8 mice. L Representative images of IHC staining analysis in Snx3-cKO mice were shown; Scale bar: 200 μm, n = 8 mice. M IF staining analysis of α-SMA were shown; Scale bar: 25 μm, n = 3 experiments. N IF staining analysis of β-catenin were shown; Scale bar: 25 μm, n = 3 experiments. The data were shown as means ± SEM. * P < 0.05 vs. CTL+ Saline group or sh-NC. # P < 0.05 vs CTL + BLM group or sh-NC + TGF-β1. & P < 0.05 vs Snx3-cKO + BLM group or sh-SNX3 + TGF-β1. ns, not significant.

Article Snippet: The following primary antibodies were used: anti-SNX3 (Proteintech, China, diluted 1:200, #10772-1-AP), anti-SNX3 (Santa Cruz, USA, diluted 1:200, #sc-376667), anti-α-SMA (Proteintech, China, diluted 1:200, #23081-1-AP), anti-ACTA2 (Proteintech, China, diluted 1:200, #14395-1-AP), anti-SP-C (Proteintech, China, diluted 1:200, #10774-1-AP), anti-Wls (Proteintech, China, diluted 1:200, #23081-1-AP), anti-CK-1α (Abcam, USA, diluted 1:200, # EPR19824 ), anti-β-catenin (Proteintech, China, diluted 1:200, #51667-2-AP), anti-PSMC3 (Cloud-Clone, China, diluted 1:200, #PAG278Hu01), anti-Rab 5A (CST, USA, diluted 1:200, #12666), and anti-LAMP1 (CST, USA, diluted 1:200, #9091).

Techniques: Co-Immunoprecipitation Assay, Immunohistochemistry, Expressing, Staining, Western Blot, Saline