FIGURE 3 from Siglec-15 Promotes Evasion of Adaptive Immunity in B-cell Acute Lymphoblastic Leukemia
Year: 2023
Type: other
Abstract: B-ALL–expressed Sig15 has dynamic subcellular localization. A, Flow cytometry of human B-ALL cell lines probed for SIG15 in both permeabilized and unpermeabilized (Surface) cells. Dotted line marks mean fluorescence intensity (MFI) of healthy PBMC signal (data representative of three independent experiments). Leukemia and PBMCs had modest surface expression of SIG15, while leukemia cells expressed higher intracellular SIG15 compared with PBMCs. SIG15 was probed using the NP159 mAb (NextCure). B and C, Immunofluorescence of REH cells staining for DAPI, Sig15, and Golgin-97. B, Imaging depicts limited surface localization of Sig15 and strong colocalization with the Golgi apparatus and TGN in unstimulated REH cells. C, Representative images depict REH cells treated for 24 hours with DMSO (vehicle control) or 81 nmol/L PMA and 1.3 μmol/L ionomycin. Stimulated REH cells show decreased colocalization of Sig15 signal with Golgin-97. SIG15 was probed using the Invitrogen polyclonal antibody. D, Flow cytometry of unpermeabilized REH cells treated with PMA with or without BOT64 for 6 hours, demonstrating that NFkB activation stimulates Sig15 localization at the cell surface (data representative of three independent experiments; dotted line represents MFI of isotype staining). SIG15 was probed using the NP159 mAb (NextCure) conjugated to Alexa Fluor 647.
Authors: Claire E. Pillsbury, Jodi Dougan, Jennifer L. Rabe, Jairo A. Fonseca, Chengjing Zhou, Alyssa N. Evans, Hasan Abukharma, Ona Ichoku, Gloria Gonzalez-Flamenco, Sunita I. Park, Ahmed Aljudi, Deborah DeRyckere, Sharon M. Castellino, Sarwish Rafiq, Solomon Langermann, Linda N. Liu, Curtis J. Henry, Christopher C. Porter
Language: English
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4384522901