Fig. 1: Viral transduction of primary human B cells from CLL/ MCL patients.

a Graph showing SLC20A1 expression analyzed by RNAseq of naive B cells (n = 55), CLL (n = 283) and MCL (n = 5) primary cells. b Schematic representation of the transduction protocol. Primary cells were co-cultured on feeder cells expressing hCD40L, hIL21 and hBAFF for 72 h prior to viral transduction through spinoculation. GFP was used as a marker for successful viral transduction. Created with BioRender.com c Transduction-efficiency as determined by GFP expression using three different envelope constructs in CLL and MCL primary cells 72 h post transduction. Representative flow cytometry images of transduction efficacy are shown for each construct on the left. Each symbol refers to an individual patient sample, n = 9 for CLL and n = 5 for MCL. Dotted line = mean ± SEM. d Quantification of viable (DAPI⁻ Annexin-V⁻), apoptotic (DAPI⁻ Annexin-V⁺) and dead (DAPI⁺) cells 3-, 5-, and 7 days post transduction (n = 8). Representative flow cytometry images and gating strategy are shown for each time point. e qRT-PCR analysis of p21 mRNA expression in primary GFP-positive CLL cells transduced with an empty vector control (EV) or a dominant-negative P53 expressing vector (P53DD) after 12 h treatment with doxorubicin, normalized to cells treated with a vehicle control (DMSO). N = 3 individual patient samples, p53WT = wild type for endogenousTP53, p53MUT = mutated endogenous TP53. The statistical significance was determined using two-tailed paired t test calculator (p = 0.0074, 0.0001 and 0.01, respectively). f Heatmap showing the percentage of apoptotic (DAPI⁻Annexin-V⁺) CLL cells transduced with either an EV or P53DD after 24 h exposure to Fludarabine. Annexin-V positivity was assessed on GFP only expressing cells. Representative flow cytometry images are shown for each drug concentration on the left (n = 4). g S-phase quantification of CLL cells transduced with an empty vector or a MYC expressing vector 6 days post transduction (n = 5). Cells co-cultured on feeder cells were pulsed with Edu for 12 h. Each symbol refers to an individual patient sample. Statistical analysis was done by a two-tailed paired t-test (p = 0.036). Cohorts are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 and not significant (ns) P > 0.05.