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. 2023 Nov 3;14:7071. doi: 10.1038/s41467-023-42820-3

Fig. 5. Dual regulation of proteins with TMP and dTAG PROTACs in OVCAR8 cells.

Fig. 5

A Schematic of genetic construct containing mCherry-FKBP12F36V and EGFP-EDHFR, with a cleavable P2A site in-between to ensure proportionally expression of each fusion protein. B OVCAR8-mCherry-FKBP12F36V-EGFP-EDHFR cells were incubated with 7c for 24 h and cell lysates were isolated for Western Blot analysis (see methods). mCherry-FKBP12F36V detected with anti-mCherry antibody and EGFP-eDHFR detected with anti-GFP antibody on the same membrane. mCherry-FKBP12F36V protein fusion is 40 kDa and eDHFR-YFP protein fusion is 45 kDa. COX IV = loading control, 18 kDa. n = 3 C OVCAR8-mCherry-FKBP12F36V-EGFP-EDHFR cells were incubated with dTAGv-1 for 24 h. mCherry-FKBP12F36V detected with anti-mCherry antibody and EGFP-eDHFR detected with anti-GFP antibody on the same membrane. mCherry-FKBP12F36V protein fusion is 40 kDa and eDHFR-YFP protein fusion is 45 kDa. COX IV = loading control, 18 kDa. n = 3 D OVCAR8-mCherry-FKBP12F36V-EGFP-EDHFR cells were incubated with 7c and dTAGv-1 for 24 h. mCherry-FKBP12F36V detected with anti-mCherry antibody and EGFP-eDHFR detected with anti-GFP antibody on the same membrane. mCherry-FKBP12F36V protein fusion is 40 kDa and eDHFR-YFP protein fusion is 45 kDa. COX IV = loading control, 18 kDa. n = 3.