MiR‐342 controls Mycobacterium tuberculosis susceptibility by modulating inflammation and cell death

A

Predicted binding sites of miR‐342‐3p and Socs6 3′‐UTR.

B

HEK‐293T fibroblasts were cotransfected with miR‐342‐3p mimic and luciferase reporter construct containing wild‐type or mutated UTR. After 24 h, cells were collected for luciferase assays. The Y‐axis showed relative luciferase activity. Data are shown as the mean ± SEM of n = 3 biological replicates.

C, D

MiR‐342‐3p mimic or inhibitor was transfected to NIH‐3T3 fibroblasts. After 48 h, cells were collected for qRT–PCR (C, data are shown as the mean ± SEM of n = 3 biological replicates) and Western blotting (D, representative blots from n = 3 biological replicates are shown) to detect SOCS6 expression.

E

€ RAW264.7 cells were transfected with Myc‐tagged Ago2 in the presence of miR‐342‐3p mimic or scramble for 24 h. Immunoprecipitation was performed with Myc antibody or IgG, and Socs6 mRNA was detected by qRT–PCR. Data are shown as the mean ± SEM of n = 3 biological replicates.

F–H

C3H BMDMs were transfected with miR‐342‐3p mimic or SOCS6‐overexpressing lentivirus, followed by stimulation with Mtb or z‐VAD (20 μM) for 24 h. Cell death mechanisms (F), cell viabilities (G), and Mtb growth rates (H) were analyzed, respectively. Data are shown as the mean ± SEM of n = 3 biological replicates.

I–K

B6 BMDMs were transfected with miR‐342‐3p inhibitor or Socs6 siRNA, followed by stimulation with Mtb or z‐VAD (20 μM) for 24 h. Cell death mechanisms (I), cell viabilities (J), and Mtb growth rates (K) were analyzed, respectively. Data are shown as the mean ± SEM of n = 3 biological replicates.

L

Relative expressions of miR‐342‐3p and Socs6 in PBMCs from healthy individuals (n = 27) and TB patients (n = 34) were detected by qRT–PCR. The correlation between miR‐342‐3p and Socs6 was analyzed by Spearman test. Spearman rank correlation: r = −0.532, P < 0.001, n = 61.

Figure 3. MicroRNA‐342‐3p directly targets SOCS6.