The 6290-bp reporter plasmid pGFPns was a gift of H. Gu (Addgene plasmid #35626) (32). The 3486-bp pmaxGFP was part of the Amaxa Nucleofection Kit (Lonza). pGFPns or pmaxGFP was first conjugated with Cy5 by Label IT Tracker Cy5 (Mirus), following the manufacturer’s protocol, with labeling density of 4 to 10 label molecules per plasmid molecule. CMPs were isolated from day 4 cultured CHRF cells, and MkMPs were purified from day 12 CD34+ cell–derived Mk culture. After washing once with 1 ml of PBS by ultracentrifugation at 25,000 rpm for 30 min at 4°C, MPs were resuspended in hypotonic buffer (Eppendorf). CMPs (5 × 106) were then mixed with Cy5-labeled pGFPns at various pGFPns/CMP ratios (5 × 103, 15 × 103, 50 × 103, 100 × 103, 250 × 103, or 500 × 103) in a total volume of 100 μl and incubated for 15 min at 37°C. pGFPns was loaded into CMPs by electroporation (Gene Pulser Xcell Electroporation System, Bio-Rad) using an exponential decay pulse for 10 to 20 ms at 100 μF, 37°C in a 2-mm cuvette. Following that, 900 μl of wash buffer (IMDM + 2% BIT + 2 mM EDTA) was added to ameliorate nucleic acid or MP aggregation, and loaded MPs were incubated on ice for 20 min for recovery, followed by centrifugation at 1000g for 10 min to remove large aggregates generated due to electroporation. Loaded MPs were collected and washed once with PBS under ultracentrifugation at 25,000 rpm, 4°C for 30 min and resuspended in PBS or coculture medium.

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