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. 2024 Sep 4;16(9):384. doi: 10.3390/toxins16090384

Figure 3.

Figure 3

Cry1JP578V competition binding assays in FAW. (A) Binding of Alexa-labeled Cry1JP578V to FAW BBMV illustrated by homologous competition. BBMVs (10 µg) were incubated with Alexa-Cry1JP578V (5 nM) in the absence (“Total”) and presence of unlabeled Cry1JP578V protein. The graphs plot the average densitometry values measured from images taken of in-gel fluorescence (see Section 5.3) for each binding reaction and normalized to the values measured in the absence of the competitor, i.e., “% Total Binding”. (B) Heterologous competition between Alexa-labeled Cry1JP578V and representatives of trait proteins with FAW BBMV show “% Total Binding” of Alexa-Cry1JP578V (5 nM) to FAW BBMV (10 μg) in the absence and presence of 1 μM unlabeled Cry1JP578V, Cry1Ac, Cry1Fa, Cry2A.127, and Vip3Ab (n = 3–12). (C) Reciprocal competitions (n = 3–6) for 2.5 nM Alexa-Cry1A.88 and 25 μg FAW BBMVs; 10 nM Alexa-Cry1Fa and 40 μg FAW BBMVs; 10 nM Alexa-Cry2A.127 and 40 μg FAW BBMVs; 10 nM Alexa Vip3Ab and 2 μg FAW BBMVs. All the error bars reflect the standard deviation. p-values < 0.01 are indicated by a bracket above compared columns. Statistical analysis was performed with one-way ANOVA with Tukey’s post hoc test for Panel (B) and with one-way ANOVA with Sidak’s post hoc test for Panel (C).