Hrp conjugated anti rabbit igg

Cells and heart tissues were lysed in lysis buffer containing protease inhibitors. Later on, the proteins were separated via 10% SDS-PAGE and transferred to a PVDF membrane. Next, the membranes were blocked in 5% milk for 2 h, before their incubation with the anti-Yap (Affinity Biosciences, cat. no. DF3182; 1:1000; China), anti-Ki67 (Affinity Biosciences, cat. no. AF0198; 1:2000; China), anti-Bax (Abclonal, cat. no. A19684; 1:2000; China), anti-Bcl2 (ABclonal, cat. no. A19693; 1:1000; China), and anti-GAPDH (Affinity Biosciences, cat. no. AF7021; 1:3000; China) at 4 °C overnight. The membranes were washed with TBST and incubated with HRP-conjugated anti-rabbit IgG (Affinity Biosciences, cat.no. S0001; 1:3000; China) for 2 h. The proteins were visualized by enhanced chemical luminescence reagents. The band intensity was analyzed with the software of Image J.

Bafilomycin A1 (Baf A1) was obtained from MedChem Express (catalog no. HY‐100558, Princeton, USA). Dorsomorphin dihydrochloride (Compound C dihydrochloride) was purchased from MedChem Express (catalog no. HY‐13418, Princeton, USA). Antibodies against p‐AMPK (1:1000; catalog no. AF3423), AMPK (1:1000; catalog no. DF6361), TFEB (1:1000; catalog no. AF7015), IL‐6 (1:1000; catalog no. DF6087) and H3 (1:3000; catalog no. BF9211) were purchased from Affinity Biosciences (OH, USA). LC3A/B (1:1000; catalog no. 4108), IL‐1β (1:1000; catalog no. 12242), GAPDH (1:1000; catalog no. 5174), p‐FOXO1/FoxO3a (1:1000; catalog no. 9464) and FOXO1 (1:1000; catalog no. 2880) were purchased from Cell Signaling Technology (Danvers, MA, USA). LAMP1 (1:1000; catalog no. A16894), ABCA1 (1:1000; catalog no. A16337), IL‐18 (1:1000; catalog no. A16737), Beclin 1 (1:1000; catalog no. A11761), CD36 (1:1000; catalog no. A5792), TNF‐α (1:1000; catalog no. A11534) and CTSD (1:1000; catalog no. A13292) were purchased from ABclonal Technology (Wuhan, China). SQSTM 1/p62 (1:1000; catalog no. ab240635) was purchased from Abcam (Cambridge, the United Kingdom). The secondary antibodies HRP‐conjugated anti‐mouse IgG and HRP‐conjugated anti‐rabbit IgG were purchased from Affinity Biosciences (OH, USA).