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. 2014 Dec 18;10(11):1921–1936. doi: 10.4161/15548627.2014.981790

Figure 7.

Figure 7.

Knockdown of Fadd reduces ER stress-induced CASP8 activation and cell death apoptosome-compromised cells upon sustained ER stress. casp9−/− MEFs stably expressing pLKO or Fadd shRNA were generated via lentiviral transduction. (A) pLKO and Fadd shRNA casp9−/− MEFs were treated with TNF alone (100 ng/ml), cycloheximide alone (1 μg/mL) or a combination of TNF and cycloheximide for 24 h after which cell viability was analyzed by propidium iodide (PI) uptake. Results are representative of at least 3 independent experiments. Error bars represent the mean ± SD. ((B)and C) casp9−/− MEFs stably expressing pLKO or Fadd shRNA were treated for the indicated times with (B) 0.5 μM Tg or (C) 0.5 μg/ml Tm, cell lysates prepared and immunoblotted for cleaved CASP8, cleaved CASP3, FADD and actin. (D and E) casp9−/− MEFs stably expressing pLKO or Fadd shRNA were treated for the indicated times with (D) 0.5 μM Tg or (E) 0.5 μg/ml of Tm and cell viability analyzed by propidium iodide (PtdIns) uptake. Results are representative of at least 3 independent experiments. Error bars represent the mean ± SD.