Skip to main content
. 2015 May 20;7(8):1004–1017. doi: 10.15252/emmm.201405010

Figure 4.

Figure 4

miR-511 is a genuine regulator of TNFR1 and has therapeutic potential

  1. Inhibition of Rluc activity of the psiCHECK-Tnfrsf1a-3′UTR reporter plasmid with B sequence (black, left panel) and S sequence (grey, right panel) by miR-511 transfection in HEK-293T cells (n = 7–8).
  2. TNFR1 protein levels in B MEF cultures (black, n = 18) and S MEF cultures (grey, n = 24) 24 h after transfection with miR-511 or miR-CTR. TNFR1 protein levels were measured in cell lysates by ELISA.
  3. TNFR1 protein levels in B MEF cultures (black, n = 18) and S MEF cultures (grey, n = 18) 24 h after transfection with anti-miR-511 or anti-miR-CTR. TNFR1 protein levels were measured in cell lysates by ELISA.
  4. In vivo effect 24 h after hydrodynamic injection of B mice with plasmids expressing miR-511 (grey, n = 19), miR-CTR (black, n = 19) or PBS (black, n = 8). Liver TNFR1 protein levels (left panel) were measured by ELISA. Twenty-four hours after plasmid injection, mice were injected with 25 μg TNF. All mice pretreated with miR-511 (grey, n = 12) survived, while 50% of the mice pretreated with miR-CTR (■, black, n = 12) or PBS (●, black, n = 12) died from TNF injection (middle panel). Twenty-four hours after injection of TNF, body temperatures of mice pretreated with miR-511 were significantly higher than those of the miR-CTR and PBS groups (right panel) (all groups n = 10).
  5. Survival of B mice and TNFR1−/− mice injected with LPS, 24 h after miR-511 or miR-CTR hydrodynamic plasmid injection. B mice pretreated with miR-511 (grey, n = 16) were significantly protected against 200 μg LPS compared to miR-CTR-pretreated mice (■, n = 15) or PBS-pretreated mice (●, n = 15) (left panel). No difference in survival was found between TNFR1−/− mice pretreated with miR-511 (grey, n = 11) or with miR-CTR (■, n = 9) against 500 μg LPS (right panel).
  6. Effect of hydrodynamic injection of plasmids expressing miR-511, miR-CTR or PBS on ConA-induced hepatitis in B mice (all groups n = 10). A total of 360 μg ConA was injected i.v. 24 h after plasmids and 8 h later mice were analysed: liver damage (serum ALT levels, left panel); body temperatures of mice 8 h after ConA injection (middle panel); TNF levels (right panel) 2 h after injection of ConA revealed similar concentrations of TNF in the three groups. miR-511-injected mice were significantly protected in the ConA model.

Data information: Data are presented as mean ± SE. Survival curves (Kaplan–Meyer plots) were compared by a log-rank test, and final outcomes were compared by a chi-square test. Differences between groups were assessed by Student’s t-test using unpaired comparisons and two-tailed analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.