Figure 4

Cln6 cortical neurons display reduced Cln6 transcripts and Zip7 staining with increased labile Zn accumulation. (A) Cln6 mRNA expression in primary murine cortical neurons was measured using qRT-PCR. Expression values were normalized to tubulin using the delta Ct method. All data are mean + SEM. (B) Zip7 expression was assessed by immunofluorescence. Zip7 was labeled with rabbit Zip7 antibodies and AlexaFluor conjugated anti-rabbit antibodies. Nuclei were stained with DAPI. Quantitative analysis of Zip7 perinuclear distribution was performed using the ArrayScan reader in conjunction with the compartmental analysis software on >1000 cells per genotype. (C) Labile Zn in control and Cln6 cortical neurons was measured by FluoZin-3 fluorescence. (D) Zip7 immunofluorescence images are representative of 3 experiments performed on triplicate coverslips. Scale bars represent 10 μm.