HG-EMMPRIN is upregulated in patients with progressive CLL. (A) EMMPRIN mRNA expression levels were quantified by qPCR in HDs and M-IGHV and UM-IGHV patients with CLL. (B) EMMPRIN protein levels in B-CLL cells of progressive and indolent patients were assessed by FC. Relative expression of percent positive cells and MFI analysis demonstrated higher levels of EMMPRIN in B-CLL cells of progressive patients than indolent patients. (C) CLL PBMC protein lysates were treated with PNGase F, and EMMPRIN expression was analyzed by western blot. GAPDH detection (bottom) was performed as loading control. The HG and LG forms are visible in untreated samples. The protein core is exposed after PNGase F treatment, confirming N-type glycosylation. (D) Representative blot of EMMPRIN in patients with progressive and indolent CLL, and relative protein expression was normalized to GAPDH. (E) Representative western blot of EMMPRIN expression in 1 patient with progressive CLL and 1 with indolent CLL after PBMC stimulation with IgM, CPG⁺IL15, and CD40L⁺IL4. (F) FC analysis of EMMPRIN expression upon stimulation with IgM, CPG⁺IL-15, and CD40L⁺IL-4 (n = 10 per group). ∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .005; ∗P < .05 panels A-E, unpaired t test; panel G, t test. Ctrl, control of unstimulated cells; LG, low glycosylated; MFI, median fluorescence intensity; NA, not activated; ns, not significant; PNGase F, peptide N-glycosidase F; WB, western blot.