Anti cd5 apc cy7

mRNA translation was analyzed using O-propargyl-puromycin (OPP)-labeling and polysome profiling, as described [22 (link)]. For analysis of CLL samples, cells were stained with anti-CD5-PerCyP5.5 and anti-CD19-pacific blue antibodies (BD Biosciences) for 15 min on ice and OPP-labeling was quantified in CD5⁺CD19⁺ cells. For PBMCs from healthy donors, cells were stained with anti-CD5-APC-Cy7, anti-CD19-pacific blue, anti-CD27-PerCP-Cy5.5 and anti-IgG-FITC antibodies (all Biolegend) for 15 min on ice, and OPP-labeling was quantified separately on CD5⁻CD19⁺IgG⁻CD27⁻ and CD5⁻CD19⁺IgG⁻CD27⁺ cells. Polysome profiling was performed as described [22 (link)].

To determine immune cell populations, the following antibodies were employed: anti-CD19-APC, anti-CD3-APC, anti-CD4-PerCP, and anti-CD8-APC (all from Immunostep); anti-CD3-FITC and anti-CD56-APC (Cytognos); and anti-CD3-PE/Cy7 and anti-CD5-APC/Cy7 (Biolegend). The subsets of immune cells were identified as follows: T cells were defined as CD3⁺CD56⁻, NK cells were identified as CD3⁻CD56⁺ and healthy B cells as CD19⁺. Leukemia cells were defined as CD19⁺, as the percentage of healthy B cells (CD19⁺CD5⁻) detected in PBMCs from patients with CLL was <2% (data not shown). Additionally, antibodies from Biolegend were used to detect surface expression of CD79A (clone: HM47), CD79B (clone: CB3-1), and IgM (clone: MHM-88). Cells were analyzed in a BD FACS Canto II flow cytometer with FACS Diva software (Beckton Dickinson).