Anti loxl2

LOXL2, YAP, and α-SMA protein expressions were analyzed using MSC-ex or BAPN-treated mouse tissue samples. Mouse tissue sections (4 µm thick) of formalin-fixed, paraffin-embedded liver specimens were deparaffinized in xylene and rehydrated in graded alcohol. Standard immunohistochemical procedures were performed on liver tissue sections using anti-LOXL2 (1:50; Santa Cruz Biotechnology, USA), anti-YAP (1:50; Bioworld, USA), or anti-α-SMA antibody (1:50; Bioworld, USA). Signals were visualized using 3, 3′-Diamino-benzidine tetrahydrochloride (Boster Biology, Wuhan, China). A brown membrane, cytoplasmic, and nuclear staining indicated a positive reaction according to different markers. The staining result of LOXL2, YAP, and α-SMA expression was determined by the percentage of positive cells by two investigators blinded to the data. Immunofluorescence staining was performed using anti-CD9 (1:100, Bioworld, USA), anti-LOXL2 (1:50), anti-YAP (1:50), or anti-α-SMA antibody (1:50). Negative controls with isotype IgG were run in parallel. Images were acquired using a laser scanning confocal microscope (Nikon, Tokyo, Japan).

Western blot was carried out as previously described [13 (link)]. The following primary antibodies were used (See also Supplementary Table): anti LOXL2 [20 (link)] 1:10,000, ER, GAPDH, Lamin, and β-Tubulin (1:500; Santa-Cruz). Anti-E-Cadherin (1:500), Vimentin (1:2000), HIF-1α (1:2000) and Fibronectin (1:1000) (abcam). Horseredish peroxidase-conjugated secondary antibodies (1:10,000; Jackson ImmunoResearch Laboratories).