Skip to main content
. 2020 Sep 17;27(9):1164–1180.e5. doi: 10.1016/j.chembiol.2020.06.013

Figure 2.

Figure 2

Characterization of HaloPROTAC-E L-AdPROM-Mediated GFP-ULK1 and FAM83D-GFP Degradation

(A and B) ARPE-19 ULK1GFP/GFP (A) or U2OS FAM83DGFP/GFP (B) FLAG-empty, FLAG-aGFP6M-Halo and FLAG-aGFP6M-HaloD106A binding mutant-expressing cells were treated with 250 nM (A) or 1 μM (B) HaloPROTAC-E for 24 h.

(C and D) ARPE-19 ULK1GFP/GFP (C) or U2OS FAM83DGFP/GFP (D) FLAG-empty and FLAG-aGFP6M-Halo-expressing cells were treated with 250 nM (C) or 1 μM (D) HaloPROTAC-E and 50 μM VHL inhibitor VH298 for 24 h.

(E and F) ARPE-19 ULK1GFP/GFP (E) or U2OS FAM83DGFP/GFP (F) FLAG-empty and FLAG-aGFP6M-Halo-expressing cells were treated with 250 nM (E) or 1 μM (F) HaloPROTAC-E and 1 μM pan-Cullin NEDDylation inhibitor MLN4924 for 24 h.

(G and H) ARPE-19 ULK1GFP/GFP (G) or U2OS FAM83DGFP/GFP (H) FLAG-empty and FLAG-aGFP6M-Halo-expressing cells were treated with 250 nM (G) or 1 μM (H) HaloPROTAC-E and 20 μM proteasome inhibitor MG132 for 24 h.

For (A)–(H), extracts were resolved by SDS-PAGE and transferred on to PVDF membranes, which were subjected to immunoblotting with indicated antibodies.