Supplementary Figure 4 from Persistence of Drug-Resistant Leukemic Stem Cells and Impaired NK Cell Immunity in CML Patients Depend on MIR300 Antiproliferative and PP2A-Activating Functions
Year: 2023
Type: supplementary-materials
Abstract: Figure S4. MIR300 anti-proliferative activity accounts for BMM-induced LSC entry into quiescence. A, Structure of 14q32 DLK1-DIO3 genomic imprinted locus hosting the MEG3-regulated human MIR300. B, MIR300 levels in 5-Aza- or DMSO-treated (24h) Ph+ cells. C, Effect of hypoxia on proliferation of CFSE+CD34+ CML-BC cells. D, left: Effect of MSC (HS-5)-derived CM on LAMA-84 proliferation expressed as fold changes of CFSE mean of fluorescence intensity (MFI)+/-SEM; middle: pro-apoptotic effect of PAD (FTY720; 2.5uM) and DMSO (control) on HS-5-cultured LAMA-84 cells; right: Effect of MSC (HS-5)-derived CM BCR-ABL1 expression (anti-ABL1) and activity anti-PY), phospho-BCR-ABL1, JAK2 expression and activity JAK2 Y1007/1008, PP2A activity (pPP2AY307 inactive form) and GRB2 used as a control (blots are representative of three independent experiments). E, Levels of C/EBPbeta and GRB2 mRNA and protein in HS-5 cells exposed to hypoxia (48h; 1% O2). F, Effect of neutralizing TGFbeta antibody (anti-TGFb Ab; 48h, 1.25 μg/ml) on MIR300 levels in CD34+ CML-BC cells. G, Effect of ectopic C/EBPalpha (MigR1-deltauORF-C/EBPalpha-HA) and C/EBPbeta (MigR1-C/EBPB-ERTAM) on MIR300 levels in K562 cells. Immunoblot shows levels of C/EBPbeta and GRB2 in normoxic and hypoxic K562 cells.
Authors: Giovannino Silvestri, Rossana Trotta, Lorenzo Stramucci, Justin J. Ellis, Jason G. Harb, Paolo Neviani, Shuzhen Wang, Ann-Kathrin Eisfeld, Christopher J. Walker, Bin Zhang, Klara Srutova, Carlo Gambacorti-Passerini, Gabriel Pineda, Catriona H. M. Jamieson, Fabio Stagno, Paolo Vigneri, Georgios Nteliopoulos, Philippa C. May, Alistair G. Reid, Ramiro Garzon, Denis-Claude Roy, Moutuaata M. Moutuou, Martin Guimond, Peter Hokland, Michael W. Deininger, Garrett Fitzgerald, Christopher Harman, Francesco Dazzi, Dragana Milojkovic, Jane F. Apperley, Guido Marcucci, Jianfei Qi, Katerina Machova Polakova, Ying Zou, Xiaoxuan Fan, Maria R. Baer, Bruno Calabretta, Danilo Perrotti
Language: English
Subfield: Hematology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361255663