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. 2022 Feb 23;16(3):3506–3521. doi: 10.1021/acsnano.1c07723

Figure 4.

Figure 4

Suppression of gene expression in plant cells mediated by sprayable peptide-based siRNA cargos. (a) Formulation of the siGFPS1/KH9-BP100 complex for YFP transgene suppression in transgenic Arabidopsis leaves. (b) YFP fluorescence in plant cells at 3 days after spraying (3 DAS) with a solution containing siGFPS1/KH9-BP100 complex. Scale bars = 50 μm. (c) Quantitative fluorescence intensity of YFP in plant cells at 3 DAS with siRNA/CPP complex. The distributions of YFP fluorescence from 15 ROIs (n = 15, 5 ROIs were collected from one leaf, 3 biological independent experiments) are shown as a box plot. Dots represent the fluorescence values. Black bars show the median values. (d) Immunoblot analysis of YFP and endogenous RubisCo Activase 1 protein (RCA1; a highly abundant intracellular plant protein) in soluble proteins extracted from Arabidopsis leaves at 3 DAS with siRNA/CPP complex. The membrane was stained with Ponceau S before probing with antibodies. RbcL = RubisCo large subunit. (e) Relative abundance of YFP/RCA1 in total leaf proteins at 3 DAS with siRNA/CPP complex determined by immunoblotting. Amounts of YFP relative to RCA1 are shown as a box plot. Magenta circles represent the distribution of data (n = 5). Black bars show median values. (f) Relative YFP transcript levels in leaves at 3 DAS with siRNA/CPP complex. Magenta dots represent the relative YFP transcript levels in five different experiments (n = 5). Different letters in the box plots indicate significant differences, as analyzed by one-way ANOVA with Tukey’s HSD test at p = 0.05.