ascl4  (Proteintech)

Journal: Communications Biology

Article Title: Astragalin promotes HSCs ferroptosis through NCOA4 mediated ferritinophagy to alleviate liver fibrosis in zebrafish and mice

doi: 10.1038/s42003-025-08421-0

Figure Lengend Snippet: A , B Western blot analyses of NCOA4 protein in HSC-T6 cells in NC or siNCOA4 group. C , D Expression of α-SMA, Desmin, and FTH by immunofluorescence staining in NC or siNCOA4 HSC-T6 cells treated with Ag, and the quantitative analysis. Scale bar = 10 or 50 μm. E HSC-T6 cells captured by TEM. The green arrows indicate the damaged mitochondria. The yellow arrows indicate the healthy mitochondria. The red arrows stand for autolysosome. F Quantitative PCR analyses of NCOA4 mRNA expression in activated JS-1 cells in NC or siNCOA4 group. G , H Western blot analyses of P62 and LC3B II protein in NC or siNCOA4 JS-1 cells treated with TGF-β1 or/and Ag, and the quantitative analysis. I and J Western blot analyses of ASCL4, XCT, and GPX4 protein in NC or siNCOA4 JS-1 cells treated with TGF-β1 or/and Ag, and the quantitative analysis. K JS-1 cells captured by TEM. L , M Expression of α-SMA by immunofluorescence staining in JS-1 cells in different groups, and the quantitative analysis. Scale bar = 20 μm. N Quantitative PCR analyses of FTH and FTL mRNA expression in JS-1 cells. All data presented as means ± SEM. n = 3–6 per group. (ns means p > 0.05, * p < 0.05, ** p < 0.01, ***p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies including α-SMA (1:400, proteintech), Desmin (HuaBio, 1:200), Collagen-1 (Abcam, 1:400), LC3B II (Abcam, 1:200), P62 (Abmart, 1:200), NCOA4 (DF4255, SAB, 1:100), GPX4 (ab125066, Abcam, 1:500), COX2 (ab283574, Abcam, 1:500), FTH (1:100, Abcam), ASCL4 (HY- P80977 , MCE, 1:100).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Journal: Communications Biology

Article Title: Astragalin promotes HSCs ferroptosis through NCOA4 mediated ferritinophagy to alleviate liver fibrosis in zebrafish and mice

doi: 10.1038/s42003-025-08421-0

Figure Lengend Snippet: A , B Immunofluorescence staining of α-SMA, TFR, DMT1, and GPX4 in HSC-T6 cells treated with Ag as well as vehicle or Fer-1, and the quantitative analysis. Scale bar = 10 μm. C , D ROS probe staining and Fe 2+ probe staining of HSC-T6 cells in different groups, and the quantitative analysis. Scale bar= 10 or 100 μm. E MDA assay of HSC-T6 cells in different groups. F , G Immunofluorescence staining of α-SMA in JS-1 cells in different groups, and the quantitative analysis. Scale bar = 20 μm. H Quantitative PCR analyses of GPX4 , COX2 and TFR mRNA expression in JS-1 cells. I , J Western blot analyses of ASCL4 and DMT1 protein in JS-1 cells, and the quantitative analysis. K , L Western blot analyses of LC3B II, P62, FTH and FTL protein in JS-1 cells, and the quantitative analysis. All data presented as means ± SEM. n = 3–5 per group. (ns means p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Primary antibodies including α-SMA (1:400, proteintech), Desmin (HuaBio, 1:200), Collagen-1 (Abcam, 1:400), LC3B II (Abcam, 1:200), P62 (Abmart, 1:200), NCOA4 (DF4255, SAB, 1:100), GPX4 (ab125066, Abcam, 1:500), COX2 (ab283574, Abcam, 1:500), FTH (1:100, Abcam), ASCL4 (HY- P80977 , MCE, 1:100).

Techniques: Immunofluorescence, Staining, Multiple Displacement Amplification, Real-time Polymerase Chain Reaction, Expressing, Western Blot