V450 anti ly6c

Leukocyte distribution and concentrations in the BALF were determined using multicolor flow cytometry. DARPins contain a His-tag allowing detection by a secondary Alexa Fluor 488 anti-His-tag antibody. The resuspended cell pellet was stained with antibodies solved in PBS supplemented with 0.5% bovine serum albumin (BSA). A master mix was prepared and 10 µl of the master mix was added per sample. The following antibodies were used to prepare the master mix: BV 510 anti-CD45.2, PE-Cy7 anti-CD19, PerCP-Cy5.5 anti-CD3, PE anti-Ly6G, APC anti-SiglecF (APC) and an Alexa Fluor 488 anti-His-tag antibody (all from Biolegend, USA), PE Texas Red anti-CD11b (Thermo Fisher, USA) and V450 anti-Ly6C (BD Biosciences, USA). Absolute cell counts in the BALF were determined using Trucount™ tubes (BD, USA) following the manufacturer’s instructions.

Pam3CSK4 and poly(dA:dT) were purchased from Invivogen. DAPI, Antibodies against β‐Actin and β‐Tubulin were purchased from Sigma‐Aldrich. Caspase‐1 p20 antibody (Clone 4B4.2.1) was obtained from Genentech, San Francisco USA. IL‐1β antibody was from R&D Systems. Antibodies against H2A, H2A.X, γ‐H2A.X, P53, and p‐P53(S15) were from Cell Signaling Technology. 53BP1 antibody was obtained from Novus biologicals. Anti‐HA, anti‐GFP, and anti‐BRCA1 antibodies were purchased from Santa Cruz Biotechnology. Pyhin1 and Ifi205b(Mnda) antibody were purchased from Mybiosource. mCherry, Alexa488‐Anti‐Sca‐1 was from Invitrogen and PE/Cy7‐Anti‐cKit, V450‐Anti‐Ly6C, FITC‐Anti‐GR1 were from BD Pharmingen.