T55546

Initially, the TNBC cells were lysed and the proteins were extracted using RIPA buffer (P0013B, Beyotime). Then, the proteins were separated through 10–15% SDS-PAGE and blotted onto 0.45 μm PVDF membranes (Millipore). Subsequently, the membranes were then underwent overnight incubation at 4 °C with primary antibodies as follows: anti-β-actin antibody (1:2000, 20536-1-AP, Proteintech), anti-SLC25A17 antibody (1:1000, A14840, Abclonal), anti-STAT3 (1:1000, #9139, CST), anti- Phospho-STAT3 (1:1000, #9145, CST), anti-JAK2 (1:1000, #3230, CST), anti-Phospho-JAK2 (1:1000, #3771, CST), anti-Ecadherin (1:20000, 20874-1-AP, Proteintech), anti-Vimentin (1:20000, 60330-1-Ig, Proteintech), anti-MMP2 (1:1000, 10373-2-AP, Proteintech), anti-MMP9 (1:1000, 10375-2-AP, Proteintech), anti- Cleaved PARP (1:1000, #5625, CST), anti-Cleaved Caspase-3 (1:1000, #9664, CST), anti-LC3 (1:1000, T55992, Abmart), anti-P62 (1:5000, T55546, Abmart), anti-Beclin-1 (1:1000, T55092, Abmart), anti-Bcl-2 (1:1000, T40056, Abmart), anti-Bax (1:1000, T40051, Abmart). Following primary antibody incubation, the membranes were exposed to secondary antibodies at room temperature for 1 h. Finally, the protein bands were visualized using the West Pico Plus Chemiluminescent Substrate (Thermo Fisher Scientific).

Total protein was extracted using a cell lysis buffer for Western blotting and immunoprecipitation (Beyotime, Shanghai, China) containing PMSF and a protease inhibitor cocktail. Proteins (30 μg/well) were separated using 10% SDS/PAGE and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The primary antibodies used in this study were against anti-HAPSTR1 (1:1000, OTl2B8; Origene), anti-HAPSTR1 (1:500, OTI2D1; Novusbio, Centennial, CO, USA), anti-LRPPRC (1:5000, 21175-1-AP; Proteintech), anti-β-actin (1:1000, 20536-1-AP; Proteintech), anti-Ub (1:1000, 10201-2-AP; Proteintech), anti-N-cadherin (1:5000, 66219-1-Ig; Proteintech), anti-Beclin1 (1:2000, T55092; Abmart, Berkeley Heights, NJ, USA), anti-P62/SQSTM1 (1:2000, T55546; Abmart), anti-ATG5 (1:2000, T55766; Abmart), anti-vimentin (1:500, WL01960; Wanleibio), anti-snail (1:500, WL01863; Wanleibio), and anti-E-Cadherin (1:500, WL01482; Wanleibio).