FIGURE 2 from Siglec-15 Promotes Evasion of Adaptive Immunity in B-cell Acute Lymphoblastic Leukemia
Year: 2023
Type: other
Abstract: Sig15 expression is regulated by NFκB activation in B-ALL. A, Western blot analysis shows upregulated SIG15 expression in REH cells stimulated for 24 hours with a PMA/ionomycin stimulation (81 nmol/L PMA and 1.3 μmol/L ionomycin) but not with 30 ng/mL of recombinant human MCSF and/or IL10. Densitometric analysis of SIG15 is quantified above. SIG15 was probed using the Invitrogen polyclonal antibody. B, qRT-PCR from REH cells treated with PMA (81 nmol/L) with or without pan-PKC inhibitor Gö 6983 (5 μmol/L) for 24 hours, showing that PKC inhibition abrogates SIG15 induction by PMA. (****, P < 0.0001). C, qRT-PCR from REH cells stimulated with PMA (81 nmol/L) and/or treated with calcineurin inhibitor CSA (1 μmol/L) or IKK-2 inhibitor BOT64 (10 μmol/L) for 24 hours, demonstrating that NFκB inhibition but not calcineurin inhibition abrogates PMA-induced SIG15 transcription (****, P < 0.0001). D, Western blot analysis shows SIG15 expression in REH cells following 24 hours of stimulation with 81 nmol/L PMA alone or in combination with 10 μmol/L of the IKK-2 inhibitor BOT64. NFκB inhibition attenuates PMA-induced upregulation of SIG15. SIG15 was probed using the 1F7 mAb (NextCure). E,SIG15 expression from the B-ALL TARGET database correlated with protein kinase C β (PRKCB; r = 0.35; P < 0.0001, Pearson correlation), calcineurin catalytic subunit alpha (PPP3CA; r = 0.42; P < 0.0001), and IL1β (IL1B; r = 0.33; P < 0.0001, Pearson correlation). F, Western blot analysis shows increased SIG15 protein in REH cells stimulated for 24 hours with increasing doses of recombinant human CD40L. Densitometric analysis of SIG15 is quantified above. SIG15 was probed using the 1F7 mAb (NextCure).
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: W4384560692