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Fig. 4 | Molecular Neurodegeneration

Fig. 4

From: MicroRNA-9 induces defective trafficking of Nav1.1 and Nav1.2 by targeting Navβ2 protein coding region in rat with chronic brain hypoperfusion

Fig. 4

MiR-9 downregulates the expression of Navβ2 proteins. a, Verification of uptake of miR-9 by NRNs after transfection, *P < 0.05 vs NC, # P < 0.05,  mean ± s.e.m, n = 3 independent RNA samples for each group. b, Effects of miR-9 on protein levels of endogenous Navβ2 in primary cultured neonatal rat neurons (NRNs), using western blot analysis. Cells were transfected with miR-9, AMO-9, miR-9 + AMO-9, or NC. mean ± s.e.m from 3 batches of cells for each group. *P < 0.05, **P < 0.01 vs NC; ## P < 0.01 vs miR-9. c, Effects of miR-9 on SCN2B in primary cultured neonatal rat neurons (NRNs) using qRT-PCR analysis. Cells were transfected with miR-9, AMO-9, miR-9 + AMO-9, or NC. mean ± s.e.m from 3 batches of cells for each group. d, Representative confocal microscope images showing primary culture hippocampus neuron stained for Tubulin (green, upper), Navβ2 (red, middle). A merged image depicting double positivity (yellow) is shown on the bottom after transfection with miR-9 mimics or/and AMO-miR-9, negative control, n = 3. e, Repression of Navβ2 by miR-9 using the miRNA-masking antisense oligodeoxynucleotides (ODNs) techniques in NRNs determined by Western blot analysis, n = 3 batches of cells for each group, mean ± s.e.m, **P < 0.01 vs NC; ## P < 0.01 vs miR-9, ODN1 (oligodeoxynucleotides, which masks the binding sites of miR-9, located in the position 336–358 of SCN2B CDS region); ODN-2 (oligodeoxynucleotides, which masks the binding sites of miR-9, located in the position of 575–597 of SCN2B CDS region)

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