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. 2022 May 31;13:840388. doi: 10.3389/fimmu.2022.840388

Figure 6.

Figure 6

Core 2 GlcNAc transferase interferes with the biosynthesis of the glycotope recognized by Siglec-7. (A) NK cell cytotoxicity assay of JVM-3 and MEC-1 cell lines. JVM-3 cells were more resistant than MEC-1 cells to NK cell cytotoxicity. Cytotoxicity assays were conducted in technical triplicate and repeated several times, with consistent results. Representative results are shown. Bars represent mean ± SD of technical triplicates. (B) Schematic representation of O-glycan biosynthesis in leukocytes. (C) Quantitative real-time polymerase chain reaction analysis of glycosyltransferases in JVM-3 and MEC-1 cells. ST6GALNAC4 expression was higher in JVM-3, whereas GCNT1 expression was higher in MEC-1 (*P < 0.05, **P < 0.01, and ***P < 0.001; Student’s t test). Bars represent mean ± SD of technical replicates (n = 3–6). (D) Effect of GCNT1 disruption in MEC-1 cells on Siglec-7–Fc binding. (E) Correlation of the GCNT1 and ST6GALNAC4 transcript levels and Siglec-7–Fc binding (in median fluorescence intensity [MFI]) to B cells from patients with chronic lymphocytic leukemia (n = 10). Association between gene expression and Siglec-7 binding was analyzed by linear regression.