|Dataset type:||Expression profiling by RT-PCR, Transcriptome or Gene expression|
|Number of samples:||128|
|Release date:||Apr 7 2017|
|Last update date:||Mar 1 2018|
|Dataset link||Mechanisms underlying FSHβ gene sensitivity to GnRH pulse frequency [092315data]|
750,000 cells were seeded on each tissue culture-treated coverslip and were grown for two days in DMEM supplemented with 10% FBS. Cells were incubated in DMEM supplemented with 2% charcoal-treated FBS and 20mM HEPES overnight before the pulse experiment. Coverslips were placed in inert coverslip racks and pulse patterns were achieved by moving racks among GnRH, wash and resting solutions in chambers maintained at 37°C in a water bath. The chamber solution in the water bath was DMEM supplemented with 2% charcoal-treated FBS and 20mM HEPES. For each condition/time point, a minimum of 3 biological replicates were collected. Cells were pulsed with a cycle period (T) of either 30 min, 1 h, 2 h or 4 h, a pulse duration of either 2.5 min, 5 min, or 10 min, and a pulse amplitude of either 0.5 nM, 1 nM, or 2 nM GnRH. 'No pulse control' samples were included. ET, elapsed time between starting point (i.e. pulse 1) and time of collection.