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. Author manuscript; available in PMC: 2018 Apr 9.
Published in final edited form as: Nat Med. 2017 Oct 9;23(11):1342–1351. doi: 10.1038/nm.4418

Figure 2. Cytoplasmic p53 links EGFR to intrinsic apoptosis.

Figure 2

(a) Immunoblot of indicated proteins in two responders (HK301 and HK336) expressing CRISPR/CAS9 protein with control guide RNA (sgCtrl) or p53 guide RNA (p53KO). (b) The % change, relative to vehicle control, in apoptotic priming as determined by cytochrome c release following dynamic BH3 profiling with BIM peptides in sgCtrl and p53KO cells treated with 1 μM erlotinib for 24 hours (mean ± s.d., n = 2). BIM was selected based on exhibiting the greatest dynamic range from tested synthetic BH3 peptides (Supplemental Fig. 4). Results are representative of two independent experiments. (c) Immunoblot of indicated proteins in HK301 sgCtrl, p53KO, p53KO + p53cyto, and p53KO + p53wt. (d) Immunofluorescence of p53 protein combined with DAPI staining to reveal protein localization in HK301 sgCtrl, p53KO + p53cyto, and p53KO + p53wt (scale bars = 20 μm). (e) Changes in indicated mRNA levels following 100 nM doxorubicin treatment for 24 hours in HK301 sgCtrl, p53KO, p53KO + p53cyto, and p53KO + p53wt. Levels were normalized to respective DMSO treated cells (mean ± s.d., n = 3). (f) Same as (b) but in HK301 sgCtrl, p53KO, p53KO + p53cyto, and p53KO + p53wt (mean ± s.d., n = 2). Results are representative of two independent experiments. (g) Same as (e) but in HK301 sgCtrl, p53KO, p53KO + p53R175H, p53KO + p53R273H, and p53KO + p53NES (mean ± s.d., n = 3). (h) Same as (b) and (f) but in HK301 sgCtrl, p53KO, p53KO + p53R175H, p53KO + p53R273H, and p53KO + p53NES (mean ± s.d., n = 2). Results are representative of two independent experiments. Comparisons were made using two-tailed unpaired Student's t-test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.