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. Author manuscript; available in PMC: 2015 Jul 30.
Published in final edited form as: Cell. 2014 Dec 18;159(7):1549–1562. doi: 10.1016/j.cell.2014.11.036

Figure 5. Disabling Caspases Downstream of Bak and Bax Triggers Type IIFN Production.

Figure 5

(A) Schematic diagram of the manipulation of intrinsic apoptosis in MEFs.

(B–D) (B) Viability of MEFs treated with ABT-737 ± 20–30 µM of Q-VD-Oph (QVD) or z-VAD.fmk (zVAD) for 24 hr, (C) bar graphs of caspase activity after 6 hr, and IFN-β in supernatant after 24 hr (n = 9 independent MEF lines). Means were compared using a two-tailed t test. See also Figure S2.

(E) Real-time qPCR analysis of Ifnb1 induction in Mcl1−/− MEFs (n = 3 independent MEF lines). Means were compared using a two-tailed t test. (F and G) (F) Viability of MEFs expressing Bims2A or Bims4E and treated with ABT-737 for 20–24 hr and (G) caspase activity after 6 hr.

(H and I) (H) Immunoblot of lysates of MEFs treated with ABT-737 (1 µM) for 4 hr and (I) bar graph of IFN-β in supernatant after 20–24 hr (n = 3 independent MEF lines per genotype). See also Figures S2 and S3.

(J) Bone marrow cellularity from WT (n = 22), Bak−/− Bax−/− (n = 8), Apaf1−/− (n = 5), Casp9−/− (n = 11), Casp3−/− (n = 13), Casp3−/− Casp7−/− (n = 7), and Bak−/− Bax−/− Casp9−/− (n = 9) bone marrow chimeras.

(K) Number of donor-derived LSK cells from WT (n = 22),Bak−/− Bax−/−(n = 8), Apaf1−/− (n = 5), Casp9−/−(n = 11), Casp3−/−(n = 13), Casp3−/− Casp7−/− (n = 7), and Bak−/− Bax−/− Casp9−/− (n = 9) bone marrow chimeras.

(L) IFN-β in serum of WT (n = 10), Casp9−/− (n = 5), Casp3−/− (n = 5), Casp3−/− Casp7−/− (n = 4), and Bak−/− Bax−/− Casp9−/− (n = 7) bone marrow chimeras. Not done, N.D.

(M) Real-time qPCR analysis of type I ISGs in bone marrow cells (n = 3–4 bone marrow chimeras per genotype). Means were compared using a two-tailed t test.

(N) Donor-CD45.2+ contribution to the peripheral blood B lymphocyte, T lymphocyte, myeloid cells, and bone marrow LSK cells of 1° and 2° recipients 16 weeks posttransplant (n = 3 donor fetal livers per genotype and 3 recipients per donor bone marrow).

Unless otherwise indicated, means were compared to WT using a one-way ANOVA with Bonferroni correction. Data represent the mean ± SEM. *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.005.