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. 2019 Aug 2;116(34):16997–17006. doi: 10.1073/pnas.1900748116

Fig. 1.

Fig. 1.

EglN3 regulation of BIM-EL protein is hydroxylation-dependent. (A) Photographs of dissected mouse SCG of the indicated genotype and age. (B) Anti–BIM-EL immunoblot of dissected SCG from 1-wk-old mice of the indicated genotype. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. (C) Anti–BIM-EL immunoblot of EglN3 primary MEFs of the indicated genotype. (D) Anti–BIM-EL immunoblot of SK-N-FI cells that were stably transduced with lentivirus encoding shRNA targeting EglN3 (shEglN3) or scramble control (shSCR). (E) Immunoblot analysis of EglN3 MEFs transduced with adenovirus (Ad) encoding WT EglN3, catalytic-dead mutant (H196A; Mut), or GFP control (Ad-GFP). (F) Anti–BIM-EL immunoblot analysis of HeLa cells transfected with siRNA targeting EglN1, EglN2, EglN3, or nontargeting control (SCR) as indicated. si, small interfering. (G) Immunoblot and crystal violet staining of U87 cells transduced with adenovirus encoding WT EglN3, catalytic dead mutant (H196A), or GFP-control. Forty-eight hours after adenovirus transduction, cells were supplemented with medium and maintained in culture for 1 additional week, followed by crystal violet staining. cl, cleaved. (H) Immunoblot of EglN3 primary MEFs with the indicated genotype upon normoxic or anoxic conditions for 16 h or treated with 1 mM DMOG or 50 μM FG0041. (I, Left) Crystal violet staining of U87 cells stably transduced with lentiviral pLKO shRNA targeting BIM (shBIM) or nontargeting control (shSCR) and subsequently transiently transduced for 48 h with adenovirus encoding EglN3 or GFP control. (I, Right) Corresponding immunoblot.