Mouse monoclonal anti il 1β

Immunofluorescence (IF) was performed as previously described^{43 (link)}. Cells grown on coverslip were fixed with 1% pFA in PBS containing 200 mM sucrose for 10 min. Cells were washed three times with 1 ml PBS and treated with 0.5% Triton-X100 in PBS at 4 °C for 5 min. Cells were again washed three times with 1 ml PBST (PBS containing 0.05% Tween 20). Cells were soaked in PBS containing 1% BSA at room temperature for 10 min and incubated for 30 min with 1:1000-diluted primary antibodies, such as mouse monoclonal anti-IL-1β (Santa Cruz Biotechnologies, E7-2-hIL1β), mouse monoclonal anti-p21 (Santa Cruz Biotechnologies, sc-817), mouse monoclonal anti-FLAG M2 (Sigma–Aldrich), rabbit polyclonal anti-Pol II (Bethyl Laboratories, A300-653A), rabbit polyclonal anti-Myc (Abcam, ab9106), and mouse monoclonal anti-Pol II (Covance, 8WG16). Cells were washed three times with 1 ml PBST and incubated with 1:1000-diluted secondary antibodies, Alexa Flour 488-conjugated anti-rabbit IgG (Thermo Fisher Scientific) and/or Cy3-conjugated anti-mouse IgG (Jackson ImmunoResearch) in PBS for 15 min. After washing cells twice with PBST, the coverslip was mounted onto slide glass using ProLong Gold antifade mountant (Thermo Fisher Scientific). Images were captured by a Zeiss Axioimager Z1 fluorescence microscope with an oil immersion objective lens (Plan Apochromat, 63 × , NA 1.4, Zeiss).