Fig. 6. AAV-hALDH1L1-Cre virus test in primary cultured system. (A) Map of AAV vectors expressing Cre recombinase under hALDH1L1 and CaMKIIa promoter. To investigate hALDH1L1 and CaMKIIa promoter activity in naive primary cultured cortical astrocytes, AAV-EF1a-DIO-mCherry virus was co-transduced. Reporter mCherry fluorescence protein can be expressed when a recombination occurs at Cre-loxP recombination sites (DIO: Double-floxed inverse Orientation) by Cre recombinase activity. AAV-CaMKIIa-Cre virus, whose promoter is known as excitatory neuron-specific, was used as a negative control. (B) Immunocytochemistry results showing mCherry expression by Cre recombinase in the condition of treatment with two viruses, AAV-hALDH1L1-Cre and AAV-EF1a-DIO-mCherry viruses. (C) Immunocytochemistry results showing no mCherry expression in negative control condition of treatment with two viruses, AAV-CaMKIIa-Cre and AAV-EF1a-DIO-mCherry viruses. Each color represents as following description. Blue indicates nuclear staining with DAPI; Green indicates astrocyte staining with Aldh1L1; Red indicates recombinase-dependent mCherry expression.
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