Loss of miR-146a activates an NF-κB-IL6-TNF signaling relay in mature cells. (A) Overlap between genes upregulated >1.2-fold in miR-146a^−/− vs WT LSK HSPCs and predicted miR-146a target genes.³⁸  (B) IPA of the 121 overlapping genes from panel A. Black points represent significantly enriched pathways (P < .1), and hematopoietic and immune-related pathways are labeled. (C-D) ELISA assay of IL6 or TNF in BM of WT, miR-146a^−/−, Nfkb1^−/−miR-146a^−/−, Il6^−/−miR-146a^−/−, and Tnf^−/−miR-146a^−/− mice (from IL6 ELISA: n = 15 WT, 13 miR-146a^−/−, 11 Nfkb1^−/−miR-146a^−/−, 10 Il6^−/−miR-146a^−/−, and 8 Tnf^−/−miR-146a^−/− mice; in TNF ELISA: n = 22 WT, 17 miR-146a^−/−, 12 Nfkb1^−/−miR-146a^−/−, 9 Il6^−/−miR-146a^−/−, and 10 Tnf^−/−miR-146a^−/− mice) P by Student t test, with Benjamini-Hochberg correction. (E-F) Quantitative reverse transcription-polymerase chain reaction analysis of Il6 or Tnf in LSK HSPCs, GM (CD11b⁺ and/or Gr-1⁺), B (CD19⁺), and T (CD3⁺) lineage cells (n = 8 WT; 9 miR-146a^−/− mice; P by t test). (G) ELISA assay of secreted IL6 in WT and miR-146a^−/− BMDM cultures (n = 4 replicates). P by linear regression. (H) ELISA assay of secreted TNF in WT and miR-146a^−/− BMDMs treated with IL6 or control (n = 4 replicates; P by linear regression).