Fig. 2: Designs of eGAL+/+-based reporting systems. | Nature Communications

Fig. 2: Designs of eGAL+/+-based reporting systems.

From: A generalizable approach for programming protease-responsive conformationally inhibited artificial transcriptional factors

Fig. 2

a Schematic illustrating the comparison between membrane-anchored GAL4 and cytosolic eGAL+/+ in reporting TEVp activity. b Fluorescence images and (c) statistical analysis of membrane-anchored GAL4 and cytosolic eGAL+/+ in reporting protease activity. Merged images of GFP (green), mCherry (red), and bright field are shown, scale bar = 10 μm. TEVp-induced fold changes of mCherry fluorescence are labeled for each comparison. d Schematic of activation of Jurkat T cells by TEVp-induced secretion of anti-CD3 scFv and anti-CD28 scFv in eGAL+/+ and TEVp co-transfected HEK293 cells. e IL2 secretion measured by ELISA (see “Methods”). f Schematic of TEVp-induced expression of n-GSDMD triggering cell pyroptosis. Percentages of (g) apoptotic cells (see “Methods”) and (h) LDH release (see “Methods”) for MCF-7 cells transfected with eGAL+/+ and TEVp. Error bars denote MMFI (arbs.units) ± SD, n = 3, individual data points represent independent triplicates performed on the same day. Significance (two-tailed, unpaired t test) is labeled above data, **p < 0.01, ****p < 0.0001. a, d, f Created in BioRender. He, J. (2025) https://BioRender.com/u3mvxcr. Source data are provided as a Source Data file.

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