Percp antimouse human cd11b

250‐300 μL of blood samples was collected for flow cytometry analysis on day 7 after AngII infusion. The following antibodies were used for staining: FITC antimouse Ly‐6C (Biolegend 128005), PerCP antimouse/human CD11b (Biolegend 101229) and PE antimouse CD45 (Biolegend 103105). Forward scatter (FSC) and side scatter (SSC) were used to gate live cells excluding red blood cells, debris and cell aggregates among total blood cells. Cells were acquired using a BD FACS Canto II (BD Biosciences) and analysed with FlowJo (Tree Star, Inc). The percentage of CD45⁺CD11b⁺ly6C^hi cells among the total leucocyte population were calculated and analysed.

Whole lungs were dissociated into single‐cell suspensions using the gentleMACS Dissociator. Red blood cell lysing buffer (Sigma‐Aldrich) was used for red cell lysis.
Lung cells were blocked with anti‐mouse CD16/32 (101319; Biolegend) and then stained with antibodies PerCP anti‐mouse/human CD11b (101229; Biolegend), Brilliant Violet 421 anti‐mouse F4/80 (123137; Biolegend), APC/Cy7 anti‐mouse CD45 (103116; Biolegend), PerCP/Cy5.5 anti‐mouse CD11c (117328; Biolegend), PE Siglec‐F (552126; BD Biosciences), PerCP/Cy5.5 anti‐mouse CD4 (100540; Biolegend), PE/Cy7 anti‐mouse CD3ε (100320; Biolegend), Brilliant Violet 421 anti‐mouse CD335 (NKp46) (137612; Biolegend), APC anti‐mouse CD8a (100712; Biolegend), Brilliant Violet 421 anti‐mouse Ly‐6G/Ly‐6C (Gr1) (108433; Biolegend) and Zombie Aqua Fixable Viability Kit (423102; Biolegend). Flow cytometric data acquisition was performed on BD FACS Canto II machine and data analysis was performed using FlowJo software.
Gating for CD45+AquaZombie−Siglec‐F+CD11c+Gr1− cells was used for AMs; CD45+AquaZombie−F4/80+CD11b+Gr1− for IMs CD45+AquaZombie−NKp46+ for natural killer (NK) cells, CD45+AquaZombie−CD3+CD8+ for CD8 T cells.