Fig 1: TINAGL1 activates HSCs by stabilizing PDGF-BB. (A) Scatter Chart of fibrosis-associated cytokines in the culture medium of Huh7.5 cells transfected with the TINAGL1 plasmid detected by a cytokine microarray assay. (B) PDGF-BB concentration quantified by ELISA in cell lysate and culture medium and by Western Blot in cell lysate of Huh7.5 cells transfected with the TINAGL1 plasmid (n = 3). (C) PDGFB mRNA level in Huh7.5 cells transfected with the TINAGL1 plasmid (n = 4). (D) Protein levels in Huh7.5 cells transfected with the TINAGL1-Flag plasmid and treated with cycloheximide (CHX), the intensity of protein was scanned by Image J (n =3). (E) Binding mode between human TINAGL1 and PDGF-BB. (F) Binding affinity between rhTINAGL1 (C-6 × His) and rhPDGF-BB proteins detected by SPR method. (G) Interaction of TINAGL1 and PDGF-BB in HEK293T cells detected by co-immunoprecipitation. (H) Co-localization of TINAGL1 (red) and PDGF-BB (green) in HEK293T cells (Scale bar: 5 μm). Nuclear (blue). Statistical analysis of co-localization of TINAGL1 and PDGF-BB fluorescence intensities was performed using ImageJ software. Data were expressed as mean ± standard deviation. P values were calculated by an unpaired two-tailed Student's t-test (B-D). ACTB, beta-actin; CHX, cycloheximide; PDGF-BB, platelet-derived growth factor BB; rhPDGF-BB, recombinant human platelet-derived growth factor BB; rhTINAGL1, recombinant human tubulointerstitial nephritis antigen-like 1; SPR, surface plasmon resonance.