Hek blue il 1β

All cell lines (HEK293T, HEK-Blue-IL-1β) are routinely tested for mycoplasma by PCR kit (ATCC, Manassas, VA) and kept at a low passage number to maintain less than 1 year since purchase, acquisition, or generation. HEK293T cells were obtained from ATCC (catalog # CRL-3216) and HEK-Blue-IL-1β cells were obtained from Invivogen (catalog # hkb-il1b) and all lines were verified by those sources and were grown in complete media containing DMEM (Gibco, Carlsbad, CA), 10% FBS, and appropriate antibiotics (Gibco, Carlsbad, CA). THP-1 cells were purchased from ATCC and grown in complete media containing RPMI (Gibco, Carlsbad, CA), 10% FBS, and 1% L-glutamine. For transient transfections, HEK293T cells were seeded the day prior to transfection in a 24-well plate (Genesee, El Cajon, CA) with 500 μl complete media. Cells were transiently transfected with 500 ng of total DNA and 1.5 μl of Transit X2 (Mirus Bio, Madison, WI) following the manufacturer’s protocol. HEK-Blue IL-1β reporter cells (Invivogen, San Diego, CA) were grown and assayed in 96-well plates (Genesee, El Cajon, CA).

All cell lines (HEK293T, HEK-Blue-IL-1β, and HaCaT) are routinely tested for mycoplasma by PCR kit (ATCC, Manassas, VA) and kept a low passage number to maintain less than one year since purchase, acquisition or generation. HEK293T cells were obtained from ATCC (catalog # CRL-3216), HEK-Blue-IL-1β cells were obtained from Invivogen (catalog # hkb-il1b) and HaCaT cells were obtained from the UC Berkeley Cell Culture Facility (https://bds.berkeley.edu/facilities/cell-culture) and all lines were verified by those sources. All cells were grown in complete media containing DMEM (Gibco, Carlsbad, CA), 10% FBS (Peak Serum, Wellington, CO), and appropriate antibiotics (Gibco, Carlsbad, CA). For transient transfections, HEK293T cells were seeded the day prior to transfection in a 24-well plate (Genesee, El Cajon, CA) with 500 µl complete media. Cells were transiently transfected with 500 ng of total DNA and 1.5 µl of Transit X2 (Mirus Bio, Madison, WI) following the manufacturer’s protocol. HEK-Blue IL-1β reporter cells (Invivogen, San Diego, CA) were grown and assayed in 96-well plates (Genesee, El Cajon, CA).