Fig. 5: Application of NBs to transfection with mRNA and pDNA of HeLa (adherent) and Jurkat (suspension) cells.

a Transfection experiments performed on HeLa cells: cells were transfected with eGFP-mRNA and eGFP-pDNA using 200-NBs at 0.1 and 0.3 μg/μL of effective nucleic acid concentration. NBs were added to the cell medium at a concentration of 1.3 × 10⁸ NBs/mL, incubated for 5 min, and irradiated at the VB threshold fluence. The transfection efficiency (i.e., % eGFP+ cells) and expression per cell (rMFI) was determined by flow cytometry 24 h post-transfection. Cell viability was determined in parallel by CellTiter-Glo assay. (i) Results for mRNA transfections. (ii) Representative confocal microscopy images of HeLa cells 24 h after mRNA transfection. (iii) Results for pDNA transfections. b Transfection experiments performed on Jurkat cells: experimental conditions were identical to HeLa’s except that the NB incubation time was 20 min. Electroporation experiments were performed using Nucleofection™ according to the manufacturer’s instructions. (i) Results for mRNA transfections. (ii) Representative confocal microscopy images of Jurkat cells 24 h after mRNA transfection, for one (×1) and two consecutive treatments (×2). (iii) The transfection yield calculated for mRNA transfections, showing for each case the percentage of non-viable cells (gray), the percentage of viable but untransfected cells (blue) and the percentage of viable and transfected cells (green). (iv) Results for pDNA transfections and (v) their corresponding transfection yield. All results are represented as mean ± SD for n = 3 biologically independent samples. Statistical significance (two-way ANOVA, with multiple comparisons) is indicated when appropriate (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).