For western blot analysis, the following antibodies were used: anti AIF, anti BAK, anti BAX, anti BCL-2, anti cIAP-2, anti Histone H1, anti-Mcl-1 and anti p53 from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA); anti BCL-XL, anti BID, anti cleaved caspase-3, 6 and 7, anti cleaved PARP, anti cytochrome c, anti COX IV, anti Endonuclease G, anti OMI/HtrA2, anti PUMA and anti Survivin from Cell Signaling Technology Inc. (Danvers, MA, USA); anti α-Tubulin from Oncogene (San Diego, CA, USA); anti caspase-9 and anti XIAP from BD Biosciences (San Jose, SA, USA); anti GAPDH from Thermo Fisher (Waltham, MA, USA); anti NOXA from Calbiochem; anti BIM and anti caspase-8 from Alexis Corp. (Lausen, Switzerland); anti SMAC/DIABLO from Millipore (Billerica, MA, USA).

Ploner C, Kofler R, Villunger A . 6A). Fig.

(A) p53‐inactive HN8 and HN12 HNSCC cells were infected with lentiviruses encoding shRNA for nontargeting control or Noxa (shNoxa2). Mukherjee N, Almeida A, Partyka KA, Lu Y, Schwan JV, Lambert K, Rogers M, Robinson WA, Robinson SE, Applegate AJ, Amato CM, Luo Y, Fujita M, Norris DA, Shellman YG. GSI also killed melanoma cell lines with low Apaf-1 levels.

Enhanced killing of melanoma cells by simultaneously targeting Mcl-1 and NOXA. It has been shown that BIM is upregulated upon DNA‐damaging agent in a p53‐independent manner (Delbridge et al., 2016; Happo et al., 2010). 2020 Mar 6;11(1):1228. doi: 10.1038/s41467-020-15051-z. J Invest Dermatol 2000; 114: 935–940.
The MEK inhibitor, PD184352, equally inactivates ERK1 and ERK2 (pERK in Fig. SB203580 and PD184352 were purchased from LC Laboratories (Woburn, MA, USA).

NOXA is a pro-apoptotic member of the BCL-2 family, which mediates induction of apoptosis via activation of mitochondria and the intrinsic apoptosis signaling pathway.1, 2 Cell death induction by cytotoxic drugs is mainly mediated by the intrinsic apoptosis signaling cascade and NOXA has an important role for apoptosis induction by cytotoxic drugs.3, 4, 5 As a so called ‘BH3-only’ protein of the BCL-2 family, upregulation of NOXA counteracts the pro-survival function of anti-apoptotic BCL-2 family members.3, 4, 5 As described for PUMA, another pro-apoptotic BH3-only member, the expression of NOXA is regulated by the transcription factor p53, but also by, for example, p38 and ERK.4, 6, 7, 8, 9 Although its important role for apoptosis induction by single drugs was extensively shown,3 the role of NOXA for drug combinations remains elusive so far. We then downregulated each transcription factor by specific shRNAs to examine which transcription factor was contributing to Noxa induction. Using p53‐inactive HNSCC cells, we find that cisplatin‐induced Noxa is mainly regulated at the transcriptional level. Luciferase activity was measured using the Dual‐Luciferase Reporter System (Promega) and normalized to the Renilla luciferase activity expressed by pRL‐SV40. Fulda S . CAS  ERK1/2- and CDK9-kinases are required to upregulate ATF4 by PG3-Oc which restores p53 transcriptomic-targets in cells without functional-p53. BA induced super-additive, synergistic apoptosis in combination with doxorubicin (doxo) in all six tumor cell lines tested (Figure 1a).

2013 Feb 15;45(4):127-37. doi: 10.1152/physiolgenomics.00160.2012. We conclude that GSI is highly effective in killing melanoma cells while sparing normal melanocytes. TP53 in hematological cancer: low incidence of mutations with significant clinical relevance. Hsu MY, Yang MH, Schnegg CI, Hwang S, Ryu B, Alani RM. (A) HN8 cells were treated with cisplatin (50 μ. Eberle J, Kurbanov BM, Hossini AM, Trefzer U, Fecker LF. For most drug combinations, underlying signaling mechanisms responsible for positive drug–drug interactions remain elusive. mL−1 penicillin G/streptomycin at 37 °C in a humidified, 5% CO2 incubator. BH3-only proteins in cell death initiation, malignant disease and anticancer therapy.

As expected, inhibition of p53 or NOXA did not affect cell death induction by BA or doxo if applied alone (Supplementary Figures S6C–F).