Mycobacterium tuberculosis subverts negative regulatory pathways in human macrophages to drive immunopathology
Fig 8
Mtb infection reduces stability of negative regulatory pathway genes and increases microRNA accumulation.
A, B, C. Mtb infection of macrophages suppresses total PIK3CD, MLST8 and MKNK1 mRNA levels at 24h after infection. D. In contrast, synthesis of newly transcribed mRNA for each gene is increased at 24h. E. Mtb infection of macrophages upregulates multiple microRNAs that are predicted to target the negative regulatory pathways analyzed at 48h. F. miR-7 targeting the 3’UTR of PIK3CD suppresses luciferase activity. Hela cells were co-transfected with a miR-7 expressing vector (miR-7) and a Renilla Luciferase construct harbouring either a PIK3CD-3’UTR fragment containing the miR-7-5p binding site (bp 268–275) (Wild Type, WT), the same PIK3CD-3’UTR construct with mutated miR-7-5p binding site (bp 268–275) (Mutant), or empty construct (Control). RLA: Relative Luciferase Activity. Mean and standard deviation are shown, p values are by Student’s t-test and data are representative of experiments performed on at least two occasions in triplicate. F. Combined results from 4 independent experiments.