Figure 2

T ₃ treatment increases the amount of RORα protein and RNA in organotypic cultures. P0 cerebellar slices kept for 7 DIV were cultured in the absence or the presence of 30 nM T₃. (A) Immunoblot analysis and quantification of RORα levels in extracts of untreated or T₃-treated cerebellar slices (*P < 0.05). (B) Left panel: fluorescence density of RORα immunolabeling was measured within each PC nucleus with MetaMorph software. Average values from multiple cells ± SEM are shown (*P < 0.05). Right panel: organotypic cultures after 10 DIV without T₃ (upper row) or with T₃ (30 nM) treatment (lower row). RORα-expressing cells were revealed by RORα immunolabeling (blue), PCs were revealed by CaBP immunolabeling (red) and both PCs and interneurons were revealed by parvalbumin immunolabeling (green). Note the presence of RORα-expressing interneurons (arrow) in the T₃ only treatment. (C) P0 organotypic cultures were cultured without T₃ (white bars) or with 30 nM T₃ (black bars) for 7 days. Levels of mRNA were determined by real time RT-PCR and standardized to 18 s rRNA The data are given relative to the mRNA level in untreated slices at the initial time of the culture (0 DIV). They were obtained from three independent cerebellar slices extracts (*P < 0.05; **P < 0.005). Error bars in (C) indicate mean ± standard deviation.