Molecules and Cells

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Fig. 2.

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Fig. 2. ARNT is not an endogenous substrate of CRL4DCAF15. (A) HEK293T cells were co-transfected with MYC-ARNT and DCAF15-HA. After 48 h, cell lysates were immunoprecipitated using MYC antibody-conjugated magnetic beads. Immune complexes were analyzed by immunoblotting with the indicated antibodies. (B) Lysates from shControl or shDCAF15 HEK293T cells were analyzed with the indicated antibodies (upper panel). qPCR validation of relative DCAF15 mRNA levels (lower panel). Data are presented as mean ± SD (n = 3). *P < 0.001. (C) HEK293T cells were transfected with pRL-TK, HRE luciferase reporter, and DCAF15 or vector control. After 24 h, transfected cells were additionally incubated under normoxia or hypoxia for 12 h. Luciferase activities were measured with Dual Luciferase Reporter Assay System. Data are presented as mean ± SD (n = 3). n.s., not significant (P > 0.1). (D) HEK293T cells were transfected with increasing amounts of DCAF15-HA. After 48 h, cell lysates were analyzed by immunoblotting with the indicated antibodies. (E) shControl or shDCAF15-containing HEK293T cells were transfected with pRL-TK and HRE luciferase reporter. After 24 h, transfected cells were incubated under normoxia or hypoxia for 12 h. Luciferase activities were measured with a Dual-Luciferase Reporter Assay System. Data are presented as mean ± SD (n = 3). n.s., not significant (P > 0.1).
Mol. Cells 2020;43:935~944 https://doi.org/10.14348/molcells.2020.0122
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