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. Author manuscript; available in PMC: 2020 Sep 4.
Published in final edited form as: Nature. 2020 Mar 4;579(7799):427–432. doi: 10.1038/s41586-020-2078-2

Fig. 2. A CRISPRi screen identifies a requirement for DELE1 in ATF4 induction.

Fig. 2

(a) Schematic of screen.

(b) Comparison of gene scores (defined in Methods) from the screen in untreated and oligomycin-treated conditions.

(c) Knockdown of DELE1 quantified by qPCR (n = 3 technical replicates).

(d) Reporter activation in cells expressing a non-targeting control sgRNA (NTC) or DELE1 sgRNAs (mean ± s.d., n = 3 culture wells).

(e) Immunoblot of endogenous ATF4. Cells were treated with 1.25 ng/mL of oligomycin for 16 hrs (Oli) or untreated (Un). Left, representative blot; Right, quantification of n = 2 blots.

(f) ATF4 reporter activation in cells transiently overexpressing (OE) DELE1-mClover and expressing sgRNA to knockdown (KD) HRI where indicated. (mean ± s.d., n = 3 culture wells).

(g) ATF4 reporter activation in cells transiently overexpressing (OE) HRI and expressing sgRNA to knockdown (KD) DELE1 where indicated. (mean ± s.d., n = 3 culture wells).

(h) DELE1-mClover expression from the AAVS1 safe-harbor locus is not sufficient to induce the ATF4 reporter in the absence of stress, but complements DELE1 knockdown upon oligomycin treatment (mean ± s.d., n = 3 culture wells).

(i) Co-localization of DELE1-mClover with the mitochondrial-targeted mRuby (Mito7-mRuby). Scale bar, 7 μm. Similar results for n = 2 culture wells.