Anti mouse cd117 c kit apc

XMU‐MP‐1 was provided by Selleck chemicals; 2′, 7′‐Di‐chlorodi‐hydrofluorescein diacetate (DCFDA) and dichloro‐fluorescein (DCF) were obtained from Sigma. Anti‐mouse Ly‐6G/Ly‐6C (Gr‐1) was abtained from BD bioscience. Anti‐mouse CD117(c‐kit)‐APC, anti‐mouse Ly‐6A/EA(Sca‐1)‐PE/Cy7, biotin‐conjugated anti‐mouse CD45R/B220, anti‐mouse CD11b, anti‐mouse Ter‐119 and APC‐Cy7‐conjugated streptavidin were obtained from eBioscience. Rabbit anti‐γ‐H2AX was obtained from Cell Signaling Technology. Rabbit anti‐NOX4 was obtained from Proteintech. FITC‐conjugated goat anti‐rabbit antibodies were obtained from Abcam Biotechnology. Cytofix/Cytoperm buffer, Perm/Wash buffer and Cytoperm Permeabilization Buffer Plus were obtained from BD Pharmingen.

Before the first 5-FU administration (day 65) and five days after every injection (days 75, 90 and 105), blood samples were extracted from the tail vein to analyze the effect of 5-FU administration on circulating hematopoietic stem cells. Blood samples were collected and prepared for immunostaining as previously described [7 (link)] and incubated for 30 minutes with following antibodies: anti-mouse Ly-6A/E (Sca-1) PE (12–5981 eBioscience), anti-mouse CD117 (c-Kit) APC (17–1171 eBioscience), mouse hematopoietic lineage eFluor 450 Cocktail (eBioscience 88–7772) and PE-Cy7 rat anti-mouse CD127 (560733 BD Biosciences). The number of hematopoietic stem cells (HSCs; lin-, Sca-1 +, c-kit +), common myeloid progenitors (CMPs; lin-, Sca-1 -, c-kit +), and common lymphoid progenitors (CLPs; Lin-, CD127+) were determined using a Gallios flow cytometer (Beckman Coulter) and the output was analyzed with Kaluza software (Beckman Coulter). The data were expressed as a percentage of the total selected events.