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. 2019 Apr 9;16(1):86–105. doi: 10.1080/15548627.2019.1598751

Figure 6.

Figure 6.

Overexpression of ULK1 alleviates PA-induced cell death in ulk1 KO cells. ulk1 KO MEF cells were transfected with vectors encoding FLAG-ULK1 and treated with PA (500 μM) for 18 h. (a) Cell viability was estimated using a Cell titer-Glo assay kit. Live cell numbers were expressed as absorbance at luminescence. (b) Immunoblot analysis of ULK1, cleaved CASP3, cleaved PARP, KEAP1, LC3B, and ACTB (loading control) in ulk1 KO MEF cells transfected with vectors encoding F-ULK1 and treated with PA (500 μM) for 18 h. (c) TUNEL analysis of cells treated as in (a). (d) ROS levels were determined using CM-H2DCFH-DA. Representative images are shown. Scale bar: 200 μm. (e) Quantification of TUNEL analysis and DCF fluorescence. Total mRNA was isolated from cells treated as described in (a) and subjected to qRT-PCR analysis for Gsta1, Srxn1 (f) and Hmox1, Nqo1 (g) mRNA expression. Data are shown as mean ± SD from 3 independent experiments. *p < 0.05 and **p < 0.01.