P jak1

Total protein was obtained from cells or tissues using a lysis buffer (Cell Signaling Technology, California, USA), and protein levels were quantified using the BCA kit (Shanghai Ze Ye Biotechnology Co., Ltd, Shanghai, China). Subsequently, approximately 30 μg of protein was loaded and separated by 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The proteins were then transferred to a polyvinylidene fluoride membrane (Millipore, Massachusetts, USA), followed by incubation with 5% bovine serum albumin and incubation with the primary antibodies against JAK1 (1:800, Invitrogen, Massachusetts, USA), p-JAK1 (1:800, Invitrogen, Massachusetts, USA), JAK2 (1:800, Invitrogen, Massachusetts, USA), p-JAK1 (1:800, Invitrogen, Massachusetts, USA), STAT3 (1:800, Invitrogen, Massachusetts, USA), p-STAT3 (1:800, Invitrogen, Massachusetts, USA), STAT5 (1:800, Invitrogen, Massachusetts, USA), p-STAT5 (1:800, Invitrogen, Massachusetts, USA), and GAPDH (1:800, Invitrogen, Massachusetts, USA). Subsequently, the membranes were incubated with a secondary antibody (1:2000; Invitrogen, Massachusetts, USA). Finally, enhanced chemiluminescence (Invitrogen, Massachusetts, USA) was used for the visualization of bands, followed by quantification using the ImageJ software (National Institutes of Health, USA) [19 (link)].

Salvia miltiorrhiza (LS; Place of Origin: Henan, China) was purchased from Shanghai Leiyunshang Pharmaceutical Co., Ltd. The HSC-T6 and LX-2 cell lines were purchased from Shanghai Fudan IBS Cell Resource Center. Phosphate buffer solution (PBS) was obtained from Chinese manufacturer Servicebio. Dulbecco’s Modified Eagle’s Medium (DMEM), Fetal bovine serum (FBS), Penicillin-Streptomycin, and trypsin (0.25%) were purchased from GIBCO (United States). NP-40 Lysis Buffer was procured from Beyotime (China). An EDTA-free protease inhibitor cocktail and PhosSTOP™ were purchased from Roche. Antibodies (Signal transducer and activator of transcription 3 [STAT3], phosphate signal transducer and activator of transcription [P-STAT3], tyrosine-protein kinase [JAK1], alpha-smooth muscle actin A [α-SMA], transforming growth factor β1 [TGFβ1], and GAPDH) were obtained from Abcam (United States). The polyclonal antibody P-JAK1 was purchased from Invitrogen. Recombinant human TGFβ1 was ordered from PeproTech. The IRDye® 800CW goat anti-Rabbit IgG secondary antibody was purchased from LI-COR Biotechnology (United States).An Annexin V-FITC apoptosis kit was purchased from Biolegend (United States). Cell Counting Kit-8 (CCK-8) was purchased from Beyotime Biotechnology Co., Ltd. (China).