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. 2020 May 21;14:132. doi: 10.3389/fncel.2020.00132

Figure 6.

Figure 6

shRNA mediated knockdown of CAMK2A suppresses the CAMK2A-BDNF pathway in microglia. (A) Quantitative RT-PCR results show a significant decrease in the mRNA levels of Camk2a in BV2 microglia following the shRNA-mediated knockdown of CAMK2A (shCAMK2A). Data represented as mean ± SD, (n = 4), Students t-test, ***p < 0.001. (B) Immunoblot shows that the shRNA mediated knockdown of CAMK2A leads to the downregulation of BDNF via the downregulation of pCREB in BV2 microglia when compared to negative control plasmid (pLKO) transfected microglial cells. (C) The histogram shows the significant downregulation of CAMK2A, pCREB, and BDNF in shCAMK2A microglia, compared to negative control plasmids (pLKO) transfected microglial cells. Immunoblot data has been quantified and normalized to β-actin. Phospho-CREB was then normalized to total CREB. Data represented as mean ± SD, (n = 3), Students t-test, ***p < 0.001; **p < 0.01; *p < 0.05. (D) Immunofluorescence analysis shows that CAMK2A (red) expression appears to be decreased in shCAMK2A microglial cells compared to negative control plasmids (pLKO) transfected microglial cells. (E) Immunofluorescence analysis shows that BDNF (red) expression appears to be decreased in shCAMK2A microglial cells compared to negative control plasmids (pLKO) transfected microglial cells. Lectin (green) used as microglial markers. Nuclei are stained with DAPI (blue), (n = 3), Scale bars = 20 μm.