Fig 1: Inhibition of HSPG2 and NF-kB reverses the effects of TNFSF13 on HSF fibrosis and inflammation. The expression of fibrosis-associated genes α-SMA, FN1, and COL1A1 in HSF after (A) transfection with si-HSPG2 or (C) co-treatment with TNFSF13 and PDTC were detected by qRT-PCR. The expression of fibrosis-associated proteins α-SMA, Fibronectin, and Collagen I in HSF after (B) transfection with si-HSPG2 or (D) co-treatment with TNFSF13 and PDTC were detected by Western blot. The expression of inflammation cytokines IL-1β, TNF-α, and TGF-β1 in HSF after (E) transfection with si-HSPG2 or (G) co-treatment with TNFSF13 and PDTC were detected by qRT-PCR. The expression of inflammation cytokines IL-1β, TNF-α, and TGF-β1 in HSF after (F) transfection with si-HSPG2 or (H) co-treatment with TNFSF13 and PDTC were detected by Western blot. *P<0.05, **P<0.01.
Fig 2: The function of TNFSF13 on the proliferation and migration of HSF. The cell viability of HSF (A) after treatment with different doses of TNFSF13 and (B) transfection with TNFSF13 siRNAs was detected by CCK-8 assay. The proliferation of HSF (C) after treatment with different doses of TNFSF13 and (D) transfection with TNFSF13 siRNAs was detected by EdU staining. The cell migration of HSF (E) after treatment with different doses of TNFSF13 and (F) transfection with TNFSF13 siRNAs was detected by Transwell assay. *P<0.05, **P<0.01.
Fig 3: Binding of TNFSF13 and HSPG2 in HSF activates NF-κB signaling pathway. (A) The interaction between TNFSF13 and HSPG2 was analyzed by genemania. (B) The interaction between TNFSF13 and HSPG2 was confirmed by CO-IP assay. The downstream signaling pathways of TNFSF13 were analyzed by String. (C/D) The protein expression and phosphorylation of p65 after treatment with different doses of TNFSF13 and transfection with TNFSF13 siRNAs was detected by Western blot. (E) The nuclear translocation of p-p65 was detected by cell immunofluorescence.
Fig 4: Inhibition of HSPG2 and NF-κB reverses the effects of TNFSF13 on HSF proliferation and migration. The cell viability of HSF after (A) transfection with si-HSPG2 or (B) co-treatment with TNFSF13 and PDTC was detected by CCK-8 assay. The proliferation of HSF after (C) transfection with si-HSPG2 or (D) co-treatment with TNFSF13 and PDTC was detected by EdU staining. The cell migration of HSF after (E) transfection with si-HSPG2 or (F) co-treatment with TNFSF13 and PDTC was detected by Transwell assay. **P<0.01.
Fig 5: The function of TNFSF13 on the fibrosis and inflammation of HSF. The expression of fibrosis-associated genes α-SMA, FN1, and COL1A1 in HSF (A) after treatment with different doses of TNFSF13 and (C) transfection with TNFSF13 siRNAs were detected by qRT-PCR. The expression of fibrosis-associated proteins α-SMA, Fibronectin, and Collagen I in HSF (B) after treatment with different doses of TNFSF13 and (D) transfection with TNFSF13 siRNAs were detected by Western blot. The expression of inflammation cytokines IL-1β, TNF-α, and TGF-β1 in HSF (E) after treatment with different doses of TNFSF13 and (G) transfection with TNFSF13 siRNAs were detected by qRT-PCR. The expression of inflammation cytokines IL-1β, TNF-α, and TGF-β1 in HSF (F) after treatment with different doses of TNFSF13 and (H) transfection with TNFSF13 siRNAs were detected by Western blot. *P<0.05, **P<0.01.
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