Figure 2

In vitro characterization and functional assessment of Itga4 KO 5TGM1-GFP cells compared to WT 5TGM1-GFP cells. (A) The WST-1 proliferation assay comparing the WT with the KO cells. (B) gene expression by RNAseq of proliferation-associated genes in 5TGM1 WT and Itga4 KO clones KO1 and KO2. (C) Correlation matrix from RNAseq data from patient myeloma cells comparing ITGA4 (arrow) to the proliferative MM signature genes, plasma cell identity markers, and common housekeeping genes. (D) Fluorescence bead count assay to quantify proliferation of Itga4 KO 5TGM1-GFP cells vs WT 5TGM1-GFP cells in the presence and absence of M2-10B4 (M2) stromal cells. The open and dense labels refer to cells cultured with and without M2-10B4 (M2). (E) Cell cycle assay comparing the Itga4 KO 5TGM1-GFP cells vs. WT 5TGM1-GFP cells in the presence and absence of M2-10B4 (M2). Data are shown as mean and SD. All cell biology panels represent the combination of three independent experiments. (*p < 0.05, **p < 0.01, ***p < 0.001; unpaired t test).