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. Author manuscript; available in PMC: 2021 Jul 5.
Published in final edited form as: Cell Rep. 2021 Jun 15;35(11):109264. doi: 10.1016/j.celrep.2021.109264

Figure 7. MYC deficiency increases proinflammatory cytokines, in part, by IRF4.

Figure 7.

(A–C) WT and MYC cKO BMDMs were stimulated with LPS (10 ng/ml) for the indicated time points. (A) The mRNA expression of Irf4 (n = 7). (B and C) The expression of IRF4 was determined by immunoblot using whole-cell lysates (B) or nuclear lysates (C). α-Tubulin or Lamin B served as a loading control. Data are representative of 3 experiments.

(D and E) WT and MYC cKO BMDMs were transfected with negative-control siRNA (NC) or siRNAs specific for IRF4 and stimulated with LPS (10 ng/ml).

(D) The expression of IRF4 was determined by immunoblot by using nuclear lysates after LPS stimulation for 3 h. Lamin B served as a loading control. Data are representative of 3 experiments.

(E) The mRNA expression of Il6, Il12b, Tnf, and Il1b after LPS stimulation for 6 h (n ≥ 4).

(F) MYC cKO BMDMs were treated with lactic acid (LA;50, 100 mM) for 0.5 h and then stimulated with LPS (10 ng/ml) for 6 h. The mRNA expression of Irf4 (relative to the Hprt housekeeping gene) was analyzed by qRT-PCR (n ≥ 4).

(G) WT and MYC cKO BMDMs were treated with LA (100 mM) for 0.5 h and then stimulated with LPS (10 ng/ml) for 6 h. The expression of IRF4 was determined by immunoblot by using nuclear lysates. Lamin B served as a loading control. Data are representative of 3 experiments.

All data are shown as mean ± SEM. *p < 0.05 by two-way ANOVA (A) or one-way ANOVA (E and F) with a post hoc Tukey test.