Extended Data Figure 1. In vitro modeling of tolerance and miR-222 induction upon prolonged LPS stimulation.

a, Schematic of experiments performed in (b). b, Expression of LPS-response genes in control BMDMs that have undergone the given treatments. 4 major expression patterns of LPS response genes in response to tolerization were noted (n=5 biologically independent samples). c, Schematic of experiments performed in (d). d, Cytokine production, measured by ELISA, by BMDMs re-stimulated with LPS overnight after pre-treatment with LPS for the given periods of time. Time points chosen for miRNA microarray analysis are highlighted in gray (n=3 biologically independent samples). e, Schematic of strategy for experiments performed in Fig. 1. f, Comparison of microarray (x-axis) and qPCR (y-axis) measurements of LPS-induced upregulation of miRNAs. Linear regression showing correlation between the two methods is plotted (n=16 miRNAs tested). g, qPCR verification of LPS-induced change in expression of 9 miRNAs (n=3 biologically independent samples). h, Expression of miR-222 after stimulation of BMDMs by anti-inflammatory and tolerance-inducing factors for the given lengths of time (n=5 biologically independent samples; Dex, Dexamethasone). i, Expression of miR-222 in response to LPS alone, or LPS after pre-treatment of BMDMs with IFNγ (n=4 biologically independent samples). For all bar and line graphs, mean +/− SEM is plotted. ** p < 0.01, * p < 0.05, + p < 0.1 as determined by 2-sided Student’s t-test for paired values.