Anti ha hrp

The antibodies used for immunoblotting, immunoprecipitation and immunohistochemistry were: PERM1 (Sigma, #HPA031711), diluted 1:1,000 for immunoblotting, 1:100 for immunohistochemistry; BNIP3 (Cell Signaling, #3769), diluted 1:1,000; TOM20 (Abcam, #ab56783), diluted 1:100 for immunohistochemistry; TOM20 (Sigma, #HPA011562), diluted 1:5,000 for immunoblotting, 1:100 for immunohistochemistry; GAPDH (Invitrogen, #AM4300), diluted 1:10,000 for immunoblotting; Ankyrin B (Invitrogen, #33-3700), diluted 1:1000 for immunoblotting, 1:100 for immunohistochemistry; MYH7 (Developmental Studies Hybridoma Bank [DSHB], BA-F8), MYH2 (DSHB, SC-71), MYH4 (DSHB, BF-F3), CD31/PECAM (BD Pharmingen, #553370), all diluted 1:100 for immunohistochemistry; anti-FLAG M2-HRP (Sigma, #A8592), anti-HA-HRP (Miltenyi, #130-091-972), Alexa Fluor 350 anti-mouse IgG2b (Life Technologies, #A-21140), Alexa Fluor 488 anti-mouse IgG1 (Life Technologies, #A-21121), Alexa Fluor 546 anti-mouse IgM (Life Technologies, #A-21045), Alexa Fluor 546 anti-rabbit (H+L) (Life Technologies, #A-11010), Alexa Fluor 488 anti-rat (Life Technologies, #A-11006), anti-mouse HRP (Sigma, #A9044) and anti-rabbit HRP (Sigma, #A0545), all diluted 1:4,000 for immunoblotting and 1:200 for immunohistochemistry.

5–10 million cells were suspended in 2X Laemmli Sample Buffer (Biorad 161–0737) and separated on a 4%–20% SDS gradient gel. After transfer to PVDF membrane, blots were blocked in 5% milk in PBS and probed with anti-HA HRP (Miltenyi 130-091-972), anti-H3 (Abcam 1791), or anti-tubulin (gift from Keith Gull) followed by anti-rabbit IgG-HRP or anti-mouse IgG-HRP.