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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Neurobiol Dis. 2019 Feb 25;127:193–209. doi: 10.1016/j.nbd.2019.02.020

Figure 1.

Figure 1.

Astrocyte-specific Ikbk2 gene deletion in hGfapcre/Ikbk2F/F mice. A. Conditional knockout of Ikbk2 gene in astrocytes was achieved through breeding mice with cyclic recombinase in control of the human glial fibrillary acidic promoter (hGfapcre) to mice that were homozygous for a floxed-Ikbk2 (Ikbk2F) gene resulting in deletion of the Ikbk2 (Ikbk2Δ). B. Ikbk2F/F and hGfapcre/Ikbk2F/F mice were genotyped via PCR for the presence of hGfapcre allele and presence for floxed (F) or wild-type (+) Ikbk2 allele. Genotyping of a C57/Bl6/J mouse is provided for reference. C. Primary cultures of astrocytes cultured from Ikbk2F/F and hGfapcre/Ikbk2F/F mice were assessed for culture purity through flow cytometric analysis for percent GLAST (astrocyte) and CD11b (microglia) expression. D. Genomic DNA from primary astrocytes, microglia, and neurons cultured from neonatal (day 1) and astrocytes cultured from adult (20-week) Ikbk2F/F and hGfapcre/Ikbk2F/F mice were analyzed via qPCR to assess the rate of Ikbk2 deletion. Percentages of Ikbk2F and Ikbk2Δ are shown in black and gray, respectively. E. Protein isolated from hGfapcre/Ikbk2F/F and Ikbk2F/F cultured astrocytes was analyzed for the amount of IKK2 via western blot. F. IKK2 protein levels were quantified using densitometric analysis with normalization to levels of β-Actin. Data are presented as mean ± SEM with Ikbk2F/F (black bars) and hGfapcre/Ikbk2F/F (grey bars; Student t-test. *** p < 0.001).