Figure 4.

Effects of ZIL on the nuclear transfection efficiency of CPP‐MC in plants. A) Schematic illustration of CPP‐MC‐mediated reporter gene (GFP or NanoLuc^TM luciferase (Nluc)) transfection of the nucleus in ZIL‐pretreated plants. B) CLSM images showing GFP expression in epidermal cells in ZIL‐pretreated A. thaliana cotyledons 24 h after transfection with CPP‐MC or naked pDNA. C, control sample transfected with the naked pDNA. Scale bars, 40 μm. Chlorophyll fluorescence and bright‐field images corresponding to the GFP fluorescent images are shown in Figure S7. C) Boxplot representation of the relative transfection efficiency of CPP‐MC based on the Nluc expression levels in ZIL‐untreated and ZIL‐pretreated A. thaliana seedlings 24 h post‐infiltration: the boxes represent the interquartile range, the lines within the boxes represent the median values, and the upper and lower whiskers represent the highest and lowest values, respectively. Statistical significance compared to the control (ZIL conc., 0 mM): ** P<0.01, **** P<0.0001 based on Dunnett's T3 test (n=30 biological replicates). D) Intensity size distributions, Z‐average diameters, and PDI of CPP‐MC‐S and CPP‐MC‐L based on DLS measurements (n=3). E) AFM height images of CPP‐MC‐S and CPP‐MC‐L. Scale bars, 200 nm. Color bars represent the height of the micelle. F) Boxplot representation of the relative transfection efficiency of CPP‐MC‐S and CPP‐MC‐L based on the Nluc expression levels in ZIL‐untreated and ZIL‐pretreated A. thaliana seedlings 24 h post‐infiltration. Statistical significance between the ZIL‐untreated and ZIL‐pretreated samples for each micelle was determined by Dunnett's T3 test (n=30 biological replicates).