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. 2020 Jan 27;295(16):5192–5203. doi: 10.1074/jbc.RA119.010699

Figure 2.

Figure 2.

Genome-wide ORF screen identifies ABCG1 as an enhancer of hypotonicity-induced calcium responses. Stimulus-induced calcium responses were measured from ORF-transfected HEK cells with the calcium FLIPR assay. A, HEK cells were transfected with 45 individual ORFs identified from the first GOF screen using the genome-wide ORF collection, and calcium responses induced by minimum hypotonic solution (320 mmol/kg stimulus and 340 mmol/kg assay solution) containing glutamate (final concentration, 100 μm) were measured. The relative ratios of calcium responses of ORF- to RFP-transfected HEK cells were calculated (n = 3). Higher responses were observed in HEK cells transfected with mGluR1, mGluR5, and two transcriptional variants (v1 and v2) of ABCG1. The dashed line indicates responses from RFP-transfected HEK cells. B and C, ABCG1-transfected HEK cells showed significantly different calcium responses to hypotonic solution (final concentration, 250 mmol/kg; Hypo.), but not to the isotonic solution (340 mmol/kg; Iso.) or that with glutamate (final concentration, 100 μm; Glut.); traces (B) and quantification of peak calcium response (ΔF/F) with error bars (C) are shown (n = 4). One-way ANOVA (F(2,9) = 63.64, p < 0.001) was used followed by Tukey's test (C). The data are means ± standard deviation. ***, p < 0.001.