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. 2021 Jul 20;22(9):e52252. doi: 10.15252/embr.202052252

Figure EV2. MiR‐342‐3p directly targets SOCS6 to regulate anti‐Mtb immunity.

Figure EV2

  • A
    Relative expressions of miR‐342‐2p target genes were analyzed by qRT–PCR. Data are shown as the mean ± SEM of n = 3 biological replicates.
  • B
    Cell death mechanisms of RAW264.7 cells transfected with siRNA, followed by Mtb infection for 36 h. Representative data (from n = 3 biological replicates) are shown as the mean ± SEM of technical replicates.
  • C, D
    MiR‐342‐3p mimic or inhibitor was transfected to RAW264.7 macrophages. 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 the relative levels of SOCS6.
  • E
    C3H BMDMs were transfected with miR‐342‐3p mimic or SOCS6‐overexpressing lentivirus, followed by Mtb infection. Cell death mechanisms were analyzed, respectively. Representative data (from n = 3 biological replicates) are shown as the mean ± SEM of technical replicates.
  • F
    B6 BMDMs were transfected with miR‐342‐3p inhibitor or Socs6 siRNA, followed by Mtb infection. Cell death mechanisms were analyzed, respectively. Representative data (from n = 3 biological replicates) are shown as the mean ± SEM of technical replicates.
  • G, H
    SOCS6‐overexpressing vector (G) or Socs6 siRNA (H) was mixed with polyethylenimine to form a complex, which was used to infect mice by tail vein injection (N/P ratio=8). Lungs were collected for transfection efficiency validation. Data are shown as the mean ± SEM of n = 3 biological replicates.
  • I
    Alveolar macrophages from mice treated with SOCS6‐overexpressing vector or Socs6 siRNA were collected to analyze cell death mechanisms. Data are shown as the mean ± SEM of n = 3 biological replicates.
  • J, K
    Secretion of cytokines TNF‐α, IL‐1, IL‐6, and CXCL15 in BMDMs obtained from Mir342 −/− B6 and Mir342 −/− B6 mice supplemented with Socs6 siRNA (J), or from Mir342 +/+ C3H and Mir342 +/+ C3H mice supplemented with SOCS6‐overexpressing vector (K), was detected by ELISA after Mtb stimulation. Data are shown as the mean ± SEM of n = 3 biological replicates.

Data information: ANOVA followed by Bonferroni post hoc test (A, C, I‐K) and two‐tailed Student t test (G, H) were used for data analysis. *P < 0.05, **P < 0.01. Abbreviations: n.s., not significant. NC, negative control.

Source data are available online for this figure.