Fig. 5: miR-34a-5p interacts with Aldolase A 3’UTR and represses Aldolase A levels in the NAc. | Nature Communications

Fig. 5: miR-34a-5p interacts with Aldolase A 3’UTR and represses Aldolase A levels in the NAc.

From: Paradoxical mTORC1-Dependent microRNA-mediated Translation Repression in the Nucleus Accumbens of Male Mice Consuming Alcohol Attenuates Glycolysis

Fig. 5

A Predicted interaction sites (in red) of miR-34a-5p sequence (top) and Aldolase A 3’UTR (middle). Mutated Aldolase 3’UTR is shown at the bottom. B Map of Aldolase A 3’UTR or mutant 3’UTR cloned into a luciferase reporter vector. C HEK293 cells were co-transfected with a reporter vector containing Aldolase A 3’UTR or mutant 3’UTR and miR-34a-5p or a negative control. Bar graph depicts average ± SD expressed as Firefly luminescence normalized to Renilla luminescence and relative to a reference control. Gray bars: Aldolase A or mutant Aldolase A. Hatched green bars: Aldolase A or mutant Aldolase A + miR-34a5p. Dotted blue bars: Aldolase A or mutant Aldolase A + miR control. **p < 0.01, ***p < 0.001, ns: non-significant. n = 4 independent experiments per group. Each data point represents an average of 3 technical replicates. Significance was determined using Two-way ANOVA followed by Tukey’s multiple comparisons test. miR x Aldolase A 3’UTR: F(2, 18) = 7.882, p = 0.0035, p = 0.0003, effect of Aldolase A 3’UTR: F(1, 18) = 3.144, p = 0.0931, Effect of miR: F(2, 18) = 34.75, p < 0.0001; Aldolase 3’UTR + miR34a-5p vs. Aldolase A 3’UTR + miR control, p < 0.0001; Aldolase A 3’UTR mutant + miR34a-5p vs. Aldolase A 3’UTR + miR34a-5p, p = 0.0053; Aldolase A 3’UTR + miR34a-5p vs. Aldolase A 3’UTR mutant + miR control, p < 0.0001. D Lenti-miR-34a-GFP infected NAc neurons. E, F miR-34a-5p or GFP was expressed in the NAc and Aldolase A protein level was evaluated by western blot analysis, quantified as a ratio of Aldolase A/GAPDH ± SEM and depicted as % of Aldolase A levels in the NAc of GFP infected mice. *p < 0.05. Water: n = 4 mice, Alcohol: n = 5 mice. Significance was determined using two-tailed Mann-Whitney test. F U = 0, p = 0.0159. Source data are provided as a Source Data file.

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