For these experiments, either an IL-1β neutralising antibody or an equivalent amount of an isotype control goat IgG antibody was administered i.p. to six-week-old female transgenic mice 24 hours prior to iFGFR1 activation.
RA SFs were pre-incubated either with mouse anti-human Id1 antibody (Abcam, Cambridge, MA, USA) or with an equivalent amount of a corresponding control antibody (non-specific mouse IgG) for two hours at 4°C.
After 5 days, inserts were transferred to a new 24-well plate, each well containing either 800 μl of a 2.5 μM solution of A2E in culture medium or culture medium with an equivalent amount of empty liposomes (controls).
The cells were cultured with 17AAG, Nutlin-3a, and cisplatin, alone or in combination, or an equivalent amount of dimethyl sulfoxide (vehicle control).
For every time point, separate groups of control mice were either left untreated or administered an equivalent amount of a non-specific isotype control antibody.
siRNA against 14-3-3 14-3-3 14-3-3valent amount of control siRNA solution, was mixed with a transfection reagent in OptiMEM and allowed to undergo complex formation for 30 min at room temperature.
For experiments with the caspase 1 inhibitor, 50 200 μM Z-YVAD-FMK (R&D Systems), 100 μM Z-DEVD-FMK (Cayman Chemical), or equivalent amount of control DMSO were added to THP-1 cells, 1 h prior to addition of pigment or controls.
