Fitc rat igg2a

The following antibodies were used for immunostaining. Anti-Arginase-1 (Santa cruz, sc-20150, 1:200), anti-BMP-2 (Abcam, ab6285, 1:100 or Bioss, bs-1012R, 1:100), anti-β-catenin (Abcam, ab2365, 1:100), anti-caspase-3 (Epitomics, 1476−1, 1:100), anti-F4/80 (Abcam, ab6640, 1:100), anti- HSP47 (Novus, NBP1-97491, 1:50), anti-K15 (Abcam, ab52816 or ThermoFisher, MA5-11344, 1:100), anti-Ki67 (ThermoFisher, RM-9106, 1:100), anti-perilipin-1 (Abcam, ab3526, 1:100), and anti-phospho-Smad1/5 (Cell signaling, 9516, 1:50).
The following antibodies were used for flow cytometry or cell sorting. Anti-CD45 (BD pharmingen, 553081, 1:200), anti-CD86 (eBioscience, 17-0862, 1:400), anti-CD206 (BioLegend, 141706, 1:40), and anti-F4/80 (eBioscience, 17-4801, 1:10). The corresponding isotype controls were APC/Cy7 Mouse IgG2a, κ (BioLegend, 400230), FITC Rat IgG2a, κ (BD pharmingen, 557228), PE Rat IgG2a, κ (BD pharmingen, 557229), and APC Rat IgG2a, κ (BD pharmingen, 551442).

To analyze the inflammatory reaction in peritoneal cavity, mice were injected i.p. with 5 µg BjV in 0.5 ml PBS. After 3 or 24 h, the animals were euthanized in a CO₂ chamber and the peritoneal cavities were washed with 5 ml PBS. Control animals were injected with 0.5 ml PBS. Total cells were counted in Malassez hemocytometric chambers. Peritoneal cells (1 × 10⁶) were incubated with fluorochrome-conjugated monoclonal antibodies specific for FcϵRI, cKIT, CD11b, F4/80, Gr-1, and Major Histocompatibility Complex (MHC)-II (BD Biosciences and BioLegend) labeled with Fluorescein isothiocyanate phycoerythrin (FITC), phycoerythrin (PE), Pacific blue, Allophycocyanin (APC), or AmCyan and acquired in a FACSCanto II (BD Biosciences) flow cytometer. FITC rat IgG2a (BD Pharmingen) and PE rat IgGk (BD Pharmingen) were used as control isotypes. The results were analyzed with FlowJo software, version 10.1 (Becton-Dickinson), mast cells (FcϵRI⁺ cKIT⁺), neutrophils (CD11b⁺ GR-1⁺), and macrophages (CD11b⁺ GR-1^low F4/80⁺ MHC-II⁺). The experiment was performed 3 times with 3 animals per group. The results were calculated by multiplying the frequency of the population (%) obtained from flow cytometry data by the total number of peritoneal live cells enumerated in each sample.