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. 2018 Nov 30;211(2):419–430. doi: 10.1534/genetics.118.301705

Figure 3.

Figure 3

The QS repressor blocks activity of QF-GR. (A) Construct and experimental schematic of the addition of QS to the dex-inducible gene expression system. The composition of constructs 1 and 2 are described in Figure 1. Construct 3 encodes for the QS repressor and a splice leader that links the mCherry open reading frame to mark expression of the array. The addition of the QS repressor restricts the activity of the QF-GR activator. Derepression of QS is achieved by the addition of quinic acid; therefore, only in the presence of both dex and quinic acid will QF-GR drive target gene expression. AD, activation domains; DBD, DNA-binding domain. (B) The mean percentage of GFP-positive animals in animals carrying the indicated transgenes. QF-GR::SL::mCherry is expressed using a ubiquitously expressed pro-1 promoter and QUAS drives a GFP reporter. pro-1p also drives the expression of the QS repressor, which restricts the activity of QF (Wei et al. 2012); derepression is achieved by the addition of quinic acid. The splice leader links mCherry expression to QS expression to mark sites in which the transgene is expressed. Error bars represent the SEM from two independent experiments. n ≥ 57 animals for each condition; * P < 0.05, t-test, as compared to both vehicle-treated (−) populations. (C) qRT-PCR measurement of relative GFP transcript levels in animals carrying the indicated transgenes. The graph presents mean, log2-fold change of expression of the indicated transgene/drug combinations relative to basal expression of vehicle-treated animals carrying the quas::GFP reporter (no QF-GR or QS transgenes). Animals exposed to 7.5 mg/ml quinic acid were pretreated for 24 hr prior to dex treatment. Error bars indicate the SEM from three biological replicates performed in triplicate. * reflects the comparison by t-test of animals treated with quinic acid for 24 hr and then vehicle or dex for 2 hr; ** represents the comparison by t-test of animals treated with quinic acid for 24 hr and then vehicle or dex for an additional 25 hr (for both * and **, P < 0.05).