Pe conjugated anti cd14

We collected samples from age-, sex-, and basic disease-matched populations, excluding C. albicans BSI patients and healthy donors with hyperthyroidism and a history of insulin injection, because these conditions may affect the expression of GLUT3. The demographic information and clinical characteristics of the subjects are listed in Table 1. PBMCs from patients and healthy controls were extracted as described above. Isolated cells were stained with PerCP-conjugated anti-CD3, PE-conjugated anti-CD14 (BioLegend) and FITC-conjugated anti-GLUT3 (R&D Systems) to detect the expression of GLUT3 in monocytes and analyze using FlowJo 10.0 software.

The cell surface markers of differentiation were evaluated by flow cytometry as described in our previous study [16 (link)]. AML cell lines or human primary AML cells were cultured in 6-well plates at a density of 1 × 10⁶ cells per well and treated with different compounds for 72 h. Subsequently, treated cells were resuspended in phosphate-buffered saline containing 1 mM EDTA and 2% FBS, and incubated with FITC-conjugated anti-CD11b or PE-conjugated anti-CD14 (BioLegend) for 30 min in the dark at room temperature. Flow cytometer data were collected by BD FACS Caliber (BD Biosciences, NJ, USA) and analyzed through FlowJo (BD Biosciences, NJ, USA).

Fc receptors were blocked with Fcγ blocker (BioLegend, San Diego, CA, USA), and the surface markers were stained with BV450-conjugated anti-CD3 (BioLegend, clone:UCHT1), FITC-conjugated anti-CD19 (BioLegend, clone: HIB19), APC/Cy7-conjugated anti-CD20 (BioLegend, clone: 2H7), PE-conjugated anti-CD14 (BioLegend, clone: ΦM P9), PE-conjugated anti-CD11c (BioLegend, clone: B-ly6), PerCP/Cy5.5-conjugated anti-CD56 (BioLegend, clone: B159), FITC-conjugated anti-CD16 (BioLegend, clone:3G8), PE-conjugated anti-CD123 (BioLegend, clone: 6H6), and APC/Cy7-conjugated anti-CD4 (BioLegend, clone: RPA-T4). After fixing and permeabilization, IGBP1 was stained with Alexa 647-conjugated anti-IGBP1 (Novus Biologicals, Centennial, CO, USA).