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. 2020 May 8;11:2286. doi: 10.1038/s41467-020-16209-5

Fig. 6. NELF controls IL-10 expression by constraining the transcription of AP-1.

Fig. 6

a RT-qPCR analysis of Il10 mRNA in WT and Nelfb KO BMDM stimulated with LPS for indicated time. Representative (left) and cumulative (LPS 0.5 h, n = 7, P = 0.0002) (right) data are shown. b IL-10 protein ELISA in WT and Nelfb KO BMDM stimulated with LPS for indicated time. Representative (left) and cumulative (LPS 6 h, n = 8, P = 0.0066) (right) data are shown. c NELF-E ChIP-seq tracks Jun and Fos in resting (−) and LPS-activated (+) (0.5 h) BMDM. d PRO-seq (sense strand) tracks for Jun and Fos in WT and Nelfb KO BMDM cultured with (+) or without (−) LPS for 0.5 h. e Cumulative RT-qPCR data showing expression level of Jun (n = 4, P = 0.0301) and Fos (n = 6, P = 0.0039) in WT and Nelfb KO BMDM treated with LPS for 0.5 h. f Immunoblot analysis of c-Fos, c-Jun, and p38 in whole cell lysates of WT and Nelfb KO BMDM treated with LPS for indicated time (left: a representative result of five replicates). For each replicate, c-Fos and c-Jun bands were quantified by densitometry at 60 min time point, normalized to internal control (p38) and expressed relative to WT ( = 1) (c-Jun P = 0.0041; c-Fos P = 0.0144). Mean + SD. g WT and Nelfb KO BMDM were pretreated with SP600125 (50 µM) or SB203580 (20 µM) for 0.5 h, where indicated, and Il10 expression following 0.5 h of LPS co-incubation was assessed by RT-qPCR (n = 5). Relative Il10 expression was normalized to the levels in WT cells that were set as 1. h Out of 29, 20 NELF− genes super-induced in Nelfb KO are potential (P < 0.001) AP-1 targets. *P < 0.05, **P < 0.01, ***P < 0.001, N.S. P > 0.05 by two-sided paired Student’s t test. n represents biologically independent experiments. Source data are provided as a Source Data file.