Illumina Genome Analyzer IIx sequencing; ChIP-Seq MyT1-HA in NS5
ChIP-Seq MyT1-HA in NS5
NS5 cells were cultured as previously described (Conti et al. 2005), using laminin as a media supplement (20ug/mL), and plated at a density of 28,000/cm2 NS5 MyT1-HA TetON cells were generated upon infection with MyT1-HA TetON-FUW lentivirus. Infected cells were expanded for several passages in NS-5 cell medium.  For induction of expression of the TetON-FUW constructs, doxycycline hyclate (DOX) (Sigma-Aldrich) was added to the culture to a final concentration of 2g/mL. NS5 cells were fixed sequentially with di(N-succimidyl) glutarate and 1% formaldehyde in phosphate buffered saline and then lysed, sonicated and immunoprecipitated, as previously described (Castro et al., 2011), with antibody anti-HA DNA libraries were prepared from 10ng of immunoprecipitated DNA according to the standard Illumina ChIP-seq protocol.
Experimental Factor: anti-HA (Abcam ab9110)
immunoprecipitate