Ab231577

Lung tissue samples of mice were exposed to 10% formalin and subsequently dehydrated and paraffin‐embedded to prepare into 4‐μm sections. For IHC staining, lung tissue section was incubated with anti‐DNAJB1 (1:50, ab231577, Abcam, Cambridge, MA, USA) and secondary antibodies after dewaxing, inactivation, and sealing using SP Kit (Solarbio, Beijing, China). Then, the sections were treated with DAB solution and hematoxylin followed by detected DNAJB1 positive cells under a microscope. For H&E staining, section was hatched with hematoxylin solution, Bluing Reagent, and Eosin Y Solution using H&E Staining Kit (Abcam). After being washed with absolute alcohol, the pathological changes of mice lung tissue section were observed under a microscope. For TUNEL staining, section was dewaxed, inactivated, and sealed, followed by hatched with Biotin labeling solution Streptavidin‐HRP solution, DAB solution, and hematoxylin using TUNEL Apoptosis Assay Kit (Beyotime, Shanghai, China). Then, the apoptosis of mice lung tissue cells was counted under a microscope.

Total proteins were extracted from PMVECs or lung tissues using RIPA lysis buffer (Beyotime). After being loaded on 10% SDS‐PAGE gel, protein was transferred onto PVDF membranes. Membranes were treated with anti‐DNAJB1 (ab231577, Abcam), anti‐Bax (ab32503, Abcam), anti‐cleaved‐caspase3 (ab2302, Abcam), anti‐Bcl‐2 (ab32124, Abcam), and anti‐β‐actin (ab8227, Abcam) with 1:1000 dilution, followed by incubated with secondary antibody (ab205718, Abcam). Then, ECL reagent (Beyotime) was employed to develop the protein bands.