Mouse N2A neuroblastoma and E14 ES cells were cultured as described in (Vance et al., 2014 (link)). The N2A cell line was
chosen because it has been used extensively as a model to study neural
differentiation in vitro (Shea et al.,
1985 ). Human neuroblastoma (SH-SY5Y) cells were grown in DMEM/F12 medium
supplemented with 10% FBS, 1% penicillin-streptomycin, and 1% L-glutamine at
37°C in a humidified atmosphere with 5% CO2. Biochemical
fractionation, ChIP and UV-RIP experiments was performed exactly as described in
Vance et al. (2014). (link) The following
antibodies were used: anti-DNMT1 (ab87656; Abcam, UK), anti-BRG1 (ab4081; Abcam),
anti-P66beta (ab76924; Abcam), anti-SIN3A (Active Motif, Belgium, 39,865), anti-CTCF
(Abcam, 70,303), anti-rabbit IgG control antibodies (Millipore, Billerica, MA) and
mouse monoclonal anti-FLAG M2 beads (Sigma–Aldrich) for FLAG-tagged POU3F3
experiments.
chosen because it has been used extensively as a model to study neural
differentiation in vitro (
1985
supplemented with 10% FBS, 1% penicillin-streptomycin, and 1% L-glutamine at
37°C in a humidified atmosphere with 5% CO2. Biochemical
fractionation, ChIP and UV-RIP experiments was performed exactly as described in
Vance et al. (2014). (link) The following
antibodies were used: anti-DNMT1 (ab87656; Abcam, UK), anti-BRG1 (ab4081; Abcam),
anti-P66beta (ab76924; Abcam), anti-SIN3A (Active Motif, Belgium, 39,865), anti-CTCF
(Abcam, 70,303), anti-rabbit IgG control antibodies (Millipore, Billerica, MA) and
mouse monoclonal anti-FLAG M2 beads (Sigma–Aldrich) for FLAG-tagged POU3F3
experiments.
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