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. 2013 Apr 23;8(4):e62040. doi: 10.1371/journal.pone.0062040

Figure 2. Effect of calcitriol addition to culture media on dopamine neurons.

Figure 2

Dose responsive increase in the number of TH immunoreactive neurons was obtained with addition of calcitriol to media. A) primary E12 control cultures and B) cultures with 10 nM calcitriol showing immunostaining with antibodies specific to tyrosine hydroxylase (red) and Tuj1 (green) counterstained with DAPI (blue). Scale bars: 20 µM. C) The percentage of dopamine neurons obtained from cultures with various concentrations of calcitriol added to media. Error bars represent SEM, ***p<0.001, **p<0.01, *p<0.05. The percentage of dopamine neurons obtained from primary E12 ventral mesencephalic cultures increased when calcitriol was added to media up to an optimum concentration of 10 nM. Dopamine neurons were also shown to be immunoreactive for other ventral midbrain DA neuron marker proteins: (D) aromatic acid decarboxylase (AADC); (E) Nurr1; (F) dopamine transporter (DAT); and (G) vesicular monoamine transporter 2 (VMAT2). Scale bars: 10 µM.