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. Author manuscript; available in PMC: 2017 Dec 13.
Published in final edited form as: Cell Rep. 2016 Dec 13;17(11):2955–2965. doi: 10.1016/j.celrep.2016.11.042

Figure 5. TMEM258 Controls ER Stress and Apoptosis in Colonic Epithelium.

Figure 5

(A) Mice were administered 2.5% DSS in drinking water for 7 days. Colon tissue was obtained at day 8 and processed for immunofluorescence imaging to detect the indicated markers. Scale bar, 25 μm.

(B and C) Complete knockout of Tmem258 was achieved by CRISPR targeting in colonic organoids derived from Tmem258+/- mice bred to stop-floxed Cas9-GFP mice. Simultaneous delivery of Cre recombinase and sgRNA targeting Tmem258 (or control) was achieved by lentiviral transduction. Viable Cas9-GFP cells were identified by immunofluorescence staining with anti-GFP antibody (B). Scale bar, 25 μm. In addition, percentages of GFP-positive cells were quantified by FACS (C). Data represent mean + SD.

(D) Sequencing and validation of insertion/deletion mutations in Tmem258 CRISPR organoids. Data represent mean and SD derived from 2 independent sgRNAs for exonic regions and 2 independent sgRNAs for intronic regions of Tmem258.

See also Figure S6.