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After PMA stimulation, cells were washed and then stimulated with MSU (100 ug/ml) for 24 h, as described [ 15].
When the cultured plates that are seeded with cells are subjected to biomechanical stimulation, cells are forced to adapt to the deformation of the culture plate.
Prior to stimulation, cells were serum starved for 2 hrs.
At the indicated time points following LPS stimulation, cells were harvested.
Following overnight stimulation, cells were transferred to tubes for flow cytometric acquisition (Dako, USA).
12 hr after serum stimulation, cells were incubated for 9 hr with 3.3 µM nocodazole.
Before stimulation, cells were serum-starved for 24 hours in medium with 1% FBS.
After stimulation, cells were fixed/permeabilized using a cytofix/cytoperm kit (BD Biosciences).
After ten minutes stimulation, cells in this muscle were able to generate Ca2+ transients with every second stimuli.
Following stimulation, cells were processed for nitrogen cavitation, subcellular fractionation, and detection of proteins of interest by western immunoblotting.
Prior to stimulation, cells were exposed to LA for one minute, rinsed and then added to the tubes for stimulation.
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