Supplementary Figure 1: Transduced monocytic cells stably express an anti-HER2 EVIR.
From: EVIR: chimeric receptors that enhance dendritic cell cross-dressing with tumor antigens
a) The top panel shows a schematic of the bidirectional lentiviral vector (LV) used to coordinately express the CtrlR or anti-HER2 EVIR, and GFP. The bottom panels illustrate representative flow cytometry dot plots showing expression of the EVIR, stained with an anti-F(ab')2 antibody, and GFP, in iBMM-CtrlR and iBMM-EVIR cells. The cells were analyzed 7 days post-transduction. One cell culture per LV type is shown; data are representative of 3 independent transduction experiments.
(b) Individual confocal images showing anti-scFv immunostaining (white), direct GFP fluorescence (green), actin fibers stained with phalloidin (magenta), and nuclear staining with DAPI (blue), in iBMM-CtrlR and anti-HER2 iBMM-EVIR cells. See Figure 1b for experimental details. Scale bar, 50 μm.
(c) Flow cytometry analysis of GFP expression in untransduced (UT) iBMMs, anti-HER2 iBMM-EVIR and iBMM-CtrlR cells. The iBMMs were passaged for 11 times during the 36 days that followed cell transduction. Data show percentage values. One cell culture per LV type is shown. The experiment was performed one time.
Numerical values for the experiments with quantitative data are presented in Supplementary Table 2.
