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. 2023 Oct 23;14(10):695. doi: 10.1038/s41419-023-06222-z

Fig. 5. PLK1-mediated TRIM26 phosphorylation increases GPX4 stability.

Fig. 5

a, b T98G (a) and U118 (b) cells transfected with control siRNA or PLK1 siRNA were subjected to IP and IB with the indicated antibodies. c, d T98G (c) and U118 (d) cells treated as indicated were subjected to IP and IB with the indicated antibodies. e PLA assay between TRIM26 and GPX4 in U118 cells treated with onvansertib (2 nM) or MLN0905 (2 nM). Scale bar: 20 μm. f, g Flag-TRIM26 WT, Flag-TRIM26 S127A or Flag-TRIM26 S127D was co-transfected with Myc-GPX4 into HEK293T cells. After treated with MG132 (20 µM) for 6 h. Cell lysates were subjected to d-IP and IB with the indicated antibodies. h Purified Flag-TRIM26 WT, Flag-TRIM26 S127A or Flag-TRIM26 S127D was incubated with GST or GST-GPX4 conjugated to beads. Pull-down samples and 5% of the input were analyzed by IB. Purified GST and GST-GPX4 were determined by CBB. i HEK293T cells transfected as indicated were subjected to d-IP with anti-Myc antibody and then analyzed by IB. j HEK293T cells treated as indicated were subjected to d-IP with anti-Myc antibody and then analyzed by IB. k HEK293T cells transfected as indicated were subjected to d-IP with anti-Myc antibody and then analyzed by IB. l Flag-TRIM26 WT, Flag-TRIM26 S127A or Flag-TRIM26 S127D was co-transfected with Myc-GPX4 into HEK293T cells. After treated with CHX for the indicated times, then cell lysates were subjected to IB. Quantification of GPX4 levels relative to β-actin is shown; student’s t test; **P < 0.01.