Fig. 3: In vivo experiments using the fluorescent reporter system.

a Schematic representation of the fluorescent reporter system. GR and GvTcR co-express proteins using cloned vectors. Each promoter (Ap, Sp) is cloned before the reporter protein. Green fluorescent protein (GFP) is expressed as a function of the presence or absence of light. It is a system in which the fluorescence level increases for an activator and decreases for an inhibitor. b Reporter system results are meant to verify the functionality of GvTcR. A bar graph indicates the fluorescent protein changes depending on isopropyl β-D-1-thiogalactopyranoside-induced protein expression. The results for the dark condition are represented by a dark gray bar, and the results for the light condition are represented by a white bar. P-values were assessed using a Student’s t test (****P < 0.001; n = 15 biologically independent samples) c Bar graph comparing fluorescence levels due to GR and GvTcR co-expression in two promoters (Ap and Sp). The upper lane represents the Ap results, and the lower lane represents the Sp results. The graph is marked according to dark and light conditions. When only GvTcR is present, dark gray is used as a differentiator; GvTcR + PR is marked in bright gray; GvTcR + GR is marked in grey; GvTcR + R69A/K141A double mutant is marked in white (****P < 0.001; n = 15, 10 biologically independent samples). d, e Comparison plot box chart for the dark and light conditions at the Ap and Sp promoter from (c) (left lane). Dark grey represents the dark condition result while white represents the light condition results. The right lane showed the differences in relative fluorescence ratio at the dark and light conditions for the four sample results. Samples are marked separately on the x-axis. n = 15 biologically independent samples. All samples with multiple n’s are labeled with an error bar. The standard error is calculated by dividing the standard deviation by the square root of number of measurements.