Figure 4.

Expression of AuNPs-PEI-pNPR1-GFP in healthy periwinkle leaves

(A) NPR1-GFP fluorescence was accumulated in healthy periwinkle mesophyll cells 2 days after infiltrating. The green fluorescence was the fluorescence of NPR1-GFP. The red fluorescence was the autofluorescence of chlorophyll. Scale bars, 10 μm.

(B) NPR1-GFP fluorescence co-localized with the nucleus stained by DAPI. The green fluorescence was the fluorescence of NPR1-GFP. The red fluorescence was the autofluorescence of chlorophyll. The blue fluorescence was the fluorescence of DAPI. Scale bars, 10 μm.

(C) NPR1-GFP fluorescence co-localized with NLS (Nuclear localization signal sequence)-mCherry fluorescence which located in the nucleus. The green fluorescence was the fluorescence of NPR1-GFP. The blue fluorescence was the autofluorescence of chlorophyll. The red fluorescence was the fluorescence of NLS-mCherry. Scale bars, 10 μm.

(D) Relative expression levels of NPR1-GFP mRNA in healthy periwinkle leaves after infiltration with AuNPs-PEI-pNPR1-GFP. Data were represented as means ± SD from three biological repeats. Statistical analyses were performed using Student’s t-test (∗∗∗, p < 0.001 and ns indicated no significant difference).

(E) Western blot for NPR1-GFP at different time points after AuNPs-PEI-pNPR1-GFP infiltration of healthy periwinkle leaves. Corresponding bands could be observed at day 1 and day 7. The molecular weight of NPR1-GFP was 93 kDa. Irrelevant or superfluous lanes had been eliminated from the blot.

(F) Confocal observation of healthy periwinkle leaves after AuNPs-PEI-pNPR1-GFP and AuNPs-PEI-pNLS-mCherry infiltration at different time points. The green fluorescence was the fluorescence of NPR1-GFP. The blue fluorescence was the autofluorescence of chlorophyll. The red fluorescence was the fluorescence of NLS-mCherry. Scale bars, 10 μm.