TLR4 engagement mimics BIK silencing. (A-B) Expression levels of mature miR-125b (A) or pre-miR125b-1 and pre-miR-125b-2 (B) in THP-1 monocytes without stimulation or after 4 hours of LPS stimulation (1 μg/mL). RNU6B and GAPDH expression was used as endogenous control for miRNA and pre-mRNA data normalization. Mean ± SD of 2 to 8 duplicates are shown. (C) Kinetics of BIK mRNA expression in THP-1 cells after LPS stimulation (1 μg/mL). RNA quantification presents fold change (FC) to time 0. Error bars represent the SD of 3 to 7 independent experiments. BIK protein levels were analyzed by immunoblot at the indicated time points. (D) Kinetics of the OCR measured in THP-1 after LPS stimulation (1 μg/mL). All data are mean ± SD (n = 6), representative of 3 independent experiments. (E) The OCR measurement on THP-1 after 12 hours of LPS stimulation. Error bars represent the mean ± SD of 2 to 3 replicates of 3 independent experiments. (F) BIK mRNA expression levels 48 hours after transfection with miR-125b antagomir and 4 hours of LPS stimulation (1 μg/mL). Data are presented as FC with THP-1 transfected with control pre-miRNA. Error bars represent the SD of 4 independent experiments. (G) The OCR measured on THP-1 after 48 hours of transfection with control miRNA (CTRL) or miR-125b antagomir (Anti-125b) (50 nM), including 12 hours of LPS treatment (1 μg/mL), normalized by number of cells. OCRs were measured in real time under basal conditions: oligomycin, ATP-synthetase-inhibited rate; FCCP, uncoupled rate; and rotenone + antimycin A, inhibited rate. Data are representative of 4 independent experiments. (H) Parameters of respiratory profiles are presented as mean ± SD for 4 independent experiments. *P < .05; **P < .01; ***P < .001. ns, Not significant.