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. 2023 Jul 3;13:10752. doi: 10.1038/s41598-023-37706-9

Figure 5.

Figure 5

Methylation of ULK1 at Arg532 attenuates its kinase activity. (a) In vitro ULK1 kinase was performed using recombinant GST-Beclin1 (1–86 aa) purified from bacteria and HA-ULK1 immunoprecipitated from HEK293T cells. (b, c) IB analysis of WCL derived from HEK293T cells co-transfected with ULK1 and Beclin 1 (b) or ATG13 (c). (d) IB analysis of WCL derived from BT-549 cells depleted of endogenous ULK1 and re-expressing doxycycline inducible ULK1-WT or ULK1-R532K. Cells were treated with 1 μg/ml doxycycline for 8 h before harvesting. Similar results were obtained in n ≥ 3 independent experiments in (ad).