Fig. 2: Integrative analysis of miRNA-mRNA and functional annotation.

a Examples of negative correlations between miRNAs and their experimentally validated targets: miR-451a-ABCB1, miR-181c-5p-BCL2, and miR-128-3p-LGALS3 (n = 51, multiple regression adjusting for sex, age and white blood cell count). The corresponding Pearson correlation coefficients (r) and nominal P are shown. Each individual is colored by its parasite density (from the lowest (blue) to the highest (red). The 95% confidence intervals of the line of best fit are shaded in gray. b Molecular and cellular functions obtained from the enrichment analysis of 456 genes targeted by the 88 miRNAs differentially expressed between before infection (BI) symptomatic parasitemia (SP) groups and/or associated with parasitemia. The horizontal bars show the level of significance. P values are calculated and adjusted using a right-tailed Fisher’s exact test and Benjamini–Hochberg method. The vertical red line represents the P significance threshold (−log (0.05)). c Annotation of subcategories of the most significantly enriched molecular and cellular function: Cell Death and Survival. P values are calculated using a right-tailed Fisher’s exact test. d Temporal expression changes of miR-16-5p (upper panel) and BCL2 (lower panel) during the course of infection in the discovery set: BI (n = 19), AP (n = 16), SP (n = 17), and AT (n = 16). Pairwise comparisons were done using a two-tailed Student’s t test (*indicates P < 0.05). e Temporal changes of absolute lymphocyte counts during the course of infection in the discovery set: BI (n = 19), AP (n = 16), SP (n = 17), and AT (n = 16). Pairwise comparisons were done using a two-tailed Student’s t test (*P < 0.001). f Experimental validation of apoptotic role of miR-16-5p using cell proliferation assays of HEK and Hela cells transfected with miR-16-5p mimic (n = 6 independent experiments) or a mimic negative control (n = 6 independent experiments). Each data point shows cell viability of mimic-miR-16-5p treated cells relative to mimic-miR-control treated cells 48 h after transfection. Statistical significance was assessed using a paired two-tailed Wilcoxon rank test (P = 0.031 for both cell lines). The box plots in d–f show the median, the 25th and 75th percentiles as box edges, and the 5th and 95th percentiles as bounds of whiskers.