Supplementary Figure S1 from Targeting CD20⁺ Aggressive B-cell Non–Hodgkin Lymphoma by Anti-CD20 CAR mRNA-Modified Expanded Natural Killer Cells <i>In Vitro</i> and in NSG Mice

<p>Supplementary Fig.S1: Anti-CD20 CAR enhances NK92 cells in-vitro cytolytic activity against CD20+ B-NHL cells by retroviral transduction. (A) Schematic representation of the anti-CD20 chimeric receptor in an RD114-pseudotyped retroviral vector. LTR indicates long terminal repeat; AMP, ampicillin resistance. (B)-(C) Expression of anti-CD20 CAR was confirmed in infected Jurkat cells by flow cytometry after staining with a goat anti-mouse (Fab)2 polyclonal antibody conjugated with biotin followed by streptavidin PE (y-axes); expression of GFP is also shown (x-axes) (B) and it was also confirmed in infected PBMC by western blot analysis with an anti-human CD3� antibody (C). (D) NK92 cells expressing anti-CD20 chimeric receptors or expression GFP only were incubated with BATDA labeled targets: CD20+ NK sensitive Ramos, CD20+ NK resistant Daudi and the CD20- pre-B-ALL cell line RS4;11 at the indicated E/ T ratios for 4 hours. Europium release assays were performed according to the manufacturer's instructions. Each data point represents the mean (SEM; n=3). Average values are reported as the mean {plus minus} SEM. p values using unpaired student t test were noted in (D).</p>