Western blotting was performed as previously described [41 (link)] using antibodies against: β-Actin (A5441, Sigma), cleaved caspase-3 (#9661, Cell Signaling, Danvers, MA, USA), cleaved caspase-9 (#9502, Cell Signaling), Mcl-1 (#559027, BD Biosciences, San Jose, CA, USA), Bax (#610983, BD Biosciences), Bid (#2002, Cell Signaling), Bcl-xL (#610212, BD Biosciences), Bim (#2819, Cell Signaling), Bcl-2 (#M0887, Agilent DAKO, Santa Clara, CA, USA), cytochrome c (sc-7159, Santa Cruz Biotechnology, Santa Cruz, CA, USA), COX IV (A21348, Invitrogen), PUMA [44 (link)], Bak (#06–536, EMD Millipore), Noxa (#OP180, EMD Millipore), p73 (A300–126A, Bethyl Laboratories, Montgomery, TX, USA), p-p73 (#4665, Cell Signaling), p-AKT (#4058, Cell Signaling), total AKT (#9272, Cell Signaling), p-ERK1/2 (#4376, Cell Signaling), total ERK1/2 (#9102, Cell Signaling), p-FoxO3A (#9464, Cell Signaling), total FoxO3A (07–702, EMD Millipore), p53 (sc-126, Santa Cruz), p-EGFR (#2234, Cell Signaling), total EGFR (#610016, BD Biosciences), KRAS (sc-30, Santa Cruz), p-Aurora A/B/C (#2914, Cell Signaling), total Aurora A (#4718, Cell Signaling), total Aurora B (#3094, Cell Signaling), and HA (#12CA5, Roche, Indianapolis, IN, USA).
Protocol detail
Comprehensive Western Blotting Analysis of Cell Signaling
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Variable analysis
independent variables
- Antibodies used for Western blotting
- Protein expression levels of β-Actin, cleaved caspase-3, cleaved caspase-9, Mcl-1, Bax, Bid, Bcl-xL, Bim, Bcl-2, cytochrome c, COX IV, PUMA, Bak, Noxa, p73, p-p73, p-AKT, total AKT, p-ERK1/2, total ERK1/2, p-FoxO3A, total FoxO3A, p53, p-EGFR, total EGFR, KRAS, p-Aurora A/B/C, total Aurora A, total Aurora B, and HA
- Western blotting protocol as previously described [41]
- Not explicitly mentioned
- Not explicitly mentioned
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