Sqstm p62

All primary antibodies were from commercial sources. With the exception of anti-GAPDH (1:10000) and anti-DYSF (1:250), all primary antibodies were utilized at a 1:1000 dilution in 1% non-fat milk and 1X TBS-T. All primary antibody incubations were over-night at 4°C with gentle rocking. Primary antibodies were: BiP (Cell Signaling Technology, 3183S), Caspase 3 (Abcam ab13847), Caspase 12 (Abcam, ab18766), CHOP (Abcam, ab11419), CALR (Cell Signaling Technology, 2891S), CKM (Abcam, ab174672), DYSF (Abcam, ab124684), FLAG (Sigma, F1804), GAPDH (Fitzgerald, 10R-G109a; 1:10000), GRP94 (Cell Signaling Technology, 2104S), MYOC (R&D Systems, AF2537; 1μg/μL), MYOC (Origene, TA323708), MYOC (Acris, AP10162PU-N), MYOM1 (Abcam, ab205618), MYOM2 (Abcam, ab93915), MYOM3 (Proteintech, 7692-I-AP), Obscurin (Millipore, ABT160), MURF1/TRIM63 (Abcam, ab172479), SQSTM/p62 (Cell Signaling Technology, 5114S), and Ubiquitin (Abcam, ab134953). All secondary antibodies were AP-conjugated and were utilized at a 1:2000 dilution in 1% non-fat milk and 1X TBS-T. All secondary antibodies were from Abcam (ab97107, ab97237, ab6722).

Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), Lipofectamine^™ 3000 transfection reagent, TRIzol reagent, LysoTracker Red and the high capacity cDNA reverse transcription kit were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Rapa and 3-MA were obtained from Sigma-Aldrich Co. (St Louis, MO, USA). CQ and BAY-11-7082 were obtained from Selleck (Houston, Texas, USA). Mouse anti-human monoclonal antibodies against PreS2/S, β-actin, and histone 3 were purchased from Abcam (C ambridge, UK). Anti-human monoclonal antibodies against LC3, Beclin-1, SQSTM/P62, NF-κB, p-NF-κB/p65, IκB, p-IκB, Vimentin, E-cadherin and Protein Disulfide Isomerase (PDI) were purchased from Cell Signaling Technology (Boston, MA, USA). The Immobilon Western Chemiluminescent HRP substrate was purchased from EMD Millipore (Billerica, MA, USA). The cell counting kit-8 was purchased from Beyotime (Shanghai, China). Alexa Fluor 488-conjugated goat anti-mouse IgG (H + L), Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L), DyLight 405-labeled goat anti-mouse IgG (H + L) and Alexa Fluor 488-conjugated goat anti-rabbit IgG (H + L) were purchased from ZSGB-BIO (Beijing, China). Propidium Iodide (PI)/RNase staining buffer was purchased from BD Biosciences (Franklin, NJ, USA).