Supplementary Figure 2: Anti-HER2 EVIR-expressing monocytic cells avidly adhere to HER2⁺ cancer cells.

(a) Flow cytometry histograms of HER2, mCh and mTq in MC38 cells transduced as indicated. One cell culture per LV type is shown; data are representative of at least 2 independent transduction experiments.

(b) Cell binding assay using Mo-EVIR/GFP (or control Mo-CtrlR/GFP) and MC38-HER2/mCh (or control MC38-mCh) cells incubated in suspension for 20 min at 1:1 ratio. The figure shows representative images of MC38 cells (mCh⁺, magenta) and monocytes (GFP⁺, green) transduced as indicated and imaged before flow cytometry; see Figure 1c for experimental details. Scale bar, 200 μm.

(c) Cell binding assay using Mo-EVIR/GFP (or control Mo-CtrlR/GFP) and MC38-HER2/mCh (or control MC38-mCh) cells incubated in suspension for 20 min at either 9:1 or 1:9 ratio. Data show the proportion of cells that appear either as single cells (green or pink, representing monocytes and MC38 cells, respectively) or in clusters (yellow, representing monocytes bound to MC38 cells), according to flow cytometry analysis. Data indicate mean values of two independent cell cultures per condition.

(d) Co-culture of iBMM-EVIR/GFP (or control iBMM-CtrlR/GFP), MC38-HER2/mTq (or control MC38-mTq), and iBMM-mCh cells under adherent conditions for 24 h before analysis. The left panel shows flow cytometry analysis of the percentage of events representing iBMMs either as single cells (green or pink) or in clusters with MC38 cells (yellow or grey); mean values of two independent cell cultures per condition are shown. The right panel shows representative flow cytometry dot plots. mTq/GFP double-positive events (clusters of iBMM-EVIR/GFP and MC38-HER2/mTq cells) fall in the doublet/aggregate area (red) based on FSC-A and FSC-H physical parameters.

Numerical values for the experiments with quantitative data are presented in Supplementary Table 2.