Rabbit anti fibronectin clone epr23110 46

Tissue sections or cultured HTFs were stained with rabbit anti-collagen I (clone E8F4L, catalog 72026, Cell Signaling Technology, Greenbelt, MD, USA), rabbit anti-fibronectin (clone EPR23110-46, catalog ab268020, Abcam, Cambridge, UK), mouse anti-STAT3 (clone 124H6, catalog 9139, Cell Signaling Technology, Greenbelt, MD, USA), rabbit anti–p-STAT3 (Tyr705) (clone D3A7, catalog 9145, Cell Signaling Technology, Greenbelt, MD, USA), and DAPI (catalog C0060, Solarbio, Beijing, China). The samples were analyzed using a fluorescence microscope (Olympus Corporation, Tokyo, Japan).

Protein samples were separated by SDS-polyacrylamide gel electrophoresis and electrotransferred onto polyvinylidene fluoride (PVDF) membranes (Merck Millipore, Burlington, MA, USA). After being blocked, the membranes were incubated with rabbit anti-collagen I (clone E8F4L, catalog 72026, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-fibronectin (clone EPR23110-46, catalog ab268020, Abcam, Cambridge, UK), rabbit anti-α-SMA (clone EPR5368, catalog ab124964, Abcam, Cambridge, UK), rabbit anti-SOCS3 (polyclonal, catalog AF8025, Beyotime, Shanghai, China), mouse anti-STAT3 (clone 124H6, catalog 9139, Cell Signaling Technology, Danvers, MA, USA), or rabbit anti–p-STAT3 (Tyr705) (clone D3A7, catalog 9145, Cell Signaling Technology, Danvers, MA, USA) overnight at 4 °C. Equal protein loading was confirmed by incubation with mouse anti–β-tubulin (clone 5G3, catalog 44032, Signalway antibody, Greenbelt, MD, USA). Horseradish peroxidase-conjugated secondary antibodies (Signalway antibody, Greenbelt, MD, USA) were incubated with the membranes at room temperature for 1 h. Finally, the bands were visualized with electrochemiluminescence (Merck Millipore, Burlington, MA, USA) and analyzed by ImageJ software 1.46r.