Single-cell transcriptome of dormant myeloma cells. (A) Experimental workflow for scRNAseq of dormant and reactivated MM cells. eGFP⁺DiD⁺ 5TGM1 cells were injected IV into BKAL mice. Hind limbs were isolated and individual eGFP⁺DiD^hi and eGFP⁺DiD^neg cells sorted into 96-well plates for scRNAseq. (B) Representative flow plot illustrating the gating of eGFP⁺DiD^hi dormant and eGFP⁺DiD^neg reactivated cell populations for cell isolation and scRNAseq. (C) Distribution of sequencing read coverage (left) and number of uniquely mapped genes (right) from across single (1C) and pooled (n = 20; 20C) dormant and reactivated cells. (D) Distribution of 92 External RNA Controls Consortium spike-in controls across dormant and reactivated single cells. (E) Distribution of gene biotypes for dormant and reactivated cells. (F) Transcript profile of dormant and reactivated single cells showing the expression level of Egfp and idiotype genes (left) and plasma cell (Prdm1, Sdc1), naive B-cell (Fcer2a, Pax5), and myeloid lineage (Myb) genes (right). (G) Alignment of the VDJ sequence from individual 5TGM1 cells. Each row represents a single cell. C, consensus sequence; D, dormant samples; NA, not annotated; ncRNA, noncoding RNA; R, reactivated samples.