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. 2022 May 6;8:253. doi: 10.1038/s41420-022-01044-y

Fig. 5. NUAK2 regulation of GPX4 is independent of its kinase activity.

Fig. 5

a Western blot of BT-549 cells transfected with cDNA encoding wild-type NUAK2 (NUAK2 OE) or kinase-dead NUAK2 (NUAK2 OEK81R) compared to control cells expressing eGFP. β-actin is the loading control (representative of n = 2 independent experiments). Quantification of normalized NUAK2 protein levels are shown in the right panel. b Relative mRNA expression of GPX4 by RT-PCR in BT-549 cells overexpressing either wild-type NUAK2 or NUAK2 K81R compared to eGFP-expressing controls (n = 2 independent experiments). Numbers above the brackets are p values from Student’s t-tests (one-sided). c Cell viability for these cells treated with the indicated dose of ML162 for 72 h. Calculated EC50 values were 94 nM (95% CI, 83−105 nM) for controls (eGFP), 18 nM (95% CI, 15–22 nM for NUAK2 OE and 26 nM (95% CI, 22-30 nM for NUAK2K81R OE (n = 3 independent experiments).