Skip to main content
. 2013 Feb 7;8(2):e55826. doi: 10.1371/journal.pone.0055826

Figure 1. Differential expression of Dnmt1, Dnmt3a, and Dnmt3b during RA induced neuronal differentiation of P19 cells.

Figure 1

A, P19 cells either left untreated (top panel) or RA treated for initial 2 days and further cultured for 4 days without RA (6 days, bottom panel) were immunostained with neuron specific β-III tubulin antibody and nuclei were stained using DAPI. B to D, Real time RT-PCR analysis of Dnmt1, Dnmt3a, and Dnmt3b during RA induced differentiation of P19 cells. Total RNA isolated from untreated P19 cells (D0) and RA treated P19 cells at different days (D2, D4, D6) was converted to cDNA and subjected to Real time quantitative PCR analysis using primers for Dnmt1, Dnmt3a, and Dnmt3b. GAPDH was used as an internal control while mRNA levels were relative to untreated P19 cells. Values represent mean of three independent experiments and error bars represent ± SEM. *P<0.05 versus untreated P19 cells. E, Western blot analysis revealed changes in the protein level of Dnmt1, Dnmt3a, and Dnmt3b at different days during P19 cell differentiation. Total cell protein from untreated P19 cells (D0) and RA treated P19 cells at different days (D2, D4, D6) was subjected to immunoblot analysis using Dnmt1, Dnmt3a, and Dnmt3b antibodies with β-Actin used as loading control. Scale bar: 20 µ m.