Research Paper Volume 16, Issue 1 pp 299—321

METTL3-dependent N6-methyladenosine modification is involved in berberine-mediated neuroprotection in ischemic stroke by enhancing the stability of NEAT1 in astrocytes

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Figure 7. Berberine exerts neuroprotective effects via METTL3 regulating the NEAT1/miR-377-3p/Nampt axis in post-OGD/R primary astrocytes. (A) Western blot to detect expression of METTL3 in primary astrocytes after OGD/R and berberine treatment. (B) A bar presenting the quantification of METTL3 in primary astrocytes. (C) RT-qPCR to analysis the ex-pression of NEAT1 in primary astrocytes after OGD/R following berberine treatment and transfection with sh-METTL3. (D) RT-qPCR to analyze the expression of miR-377-3p in astrocytes after OGD/R following berberine treatment and transfection with sh-METTL3. (E) Western blotting to analyze the expression of Nampt in astrocytes after OGD/R following berberine treatment and transfection with sh-METTL3. (F) A bar presenting the quantification of Nampt in primary astrocytes. (G) Using CCK-8 assay to assess the effect on co-cultured neurons by treating with sh-METTL3 and berberine-treated primary astrocytes and OGD/R treatment. (H) Using LDH assay to evaluate the effect on co-cultured neurons by treating with sh-METTL3 and berberine-treated primary astrocytes and OGD/R treatment. The relative expression levels were quantified by normalizing to β-actin. Data are represented as mean ± SD, (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001).