Ab32575

The western blot was performed as described previously [32 (link)]. The experiment was replicated three times in the study. Original bands were presented in Supplemental Material.
Primary antibodies: antibody against UHRF1(Santa Cruz, sc-373750); antibody against GPX4 (Abcam, ab125066); antibody against FSP1(ABclonal, A12128); antibody against Fibronectin (Abcam, ab45688,); antibody against Collagen I (ABclonal, A1352); antibody against α-SMA (Abcam, ab32575); antibody against DNMT1 (Santa Cruz, sc-271729); antibody against UHRF1 (Santa Cruz, H-8, sc-373750); antibody against GAPDH (ABclonal, AC002).

LX-2 cells were seeded on Poly-lysine-pre-coated glass cover slips and incubated with THP-1 cells stably over-expressing lncRNA-HEIM or mock control in transwell (3.0 μm, Corning Costar, USA) for 36 h. After washing with PBS, cells were sequentially fixed with 4% ice-cold paraformaldehyde for 30 min, permeabilized with 1% Triton X-100 for 20 min, and blocked using 5% bovine serum albumin (BSA) in PBS for 1 h at room temperature. Next, cells were incubated with primary antibodies against α-SMA (1:100, rabbit, Abcam, ab32575), COL1A2 (1:100, mouse, Santa Cruze, sc-166865) overnight at 4°C, followed by incubation with FITC-conjugated secondary antibodies (1:150, Thermo Fisher, USA) and PE-conjugated secondary antibodies (1:150, Thermo Fisher, USA) in PBS away from light for 30 min at room temperature. And the nuclei were stained with DAPI for 20 min. All immunofluorescence was then visualized using confocal microscope (NIKON A1R).

Lung tissues and cultured cells were homogenized in T-PER Tissue Protein Extraction Reagent (Thermo Scientific), and RIPA lysis buffer was added with phenylmethylsulfonyl fluoride for extraction of total proteins (Beyotime Institute of Biotechnology, Shanghai, China; PMSF, Sigma-Aldrich). A total of 80 μg of protein extracts were separated on 10% polyacrylamide gels (Sigma-Aldrich) and transferred onto polyvinylidene difluoride membranes (0.2 μm, Immobilon, ISEQ00010). Protein bands were next incubated with indicated primary antibodies for analysis of protein levels as described previously (37 (link)).
The following primary antibodies were used: antibody against fibronectin (Abcam, ab45688); antibody against collagen I (ABclonal, A1352); antibody against α-SMA (Abcam, ab32575); antibody against UHRF1 (Santa Cruz, H-8, sc-373750); antibody against beclin 1 (ABclonal, A7353); antibody against Dnmt1 (Santa Cruz, sc-271729); antibody against TEAD1 (ABclonal, A13366); antibody against TEAD4 (ABclonal, A4151); antibody against YAP (Proteintech, 13584-1-AP); and antibody against GAPDH (ABclonal, AC002).