Fig. 2: Construction and characterization of Apt-PRCs ZAJ4a-7D12. | Nature Communications

Fig. 2: Construction and characterization of Apt-PRCs ZAJ4a-7D12.

From: Engineering aptamer-directed phosphatase recruiting chimeras: a strategy for modulating receptor function and overcoming drug resistance

Fig. 2

a PAGE analysis of the constructed Apt-PRCs ZAJ4a-7D12. Lane 1, ZAJ4a-Cy3; lane 2, 7D12; lane 3, ZAJ4a-Cy3-7D12; lane 4, protein makers. Cy3 was conjugated at the 5’ end of ZAJ4a. Left, Coomassie bright blue staining; right, fluorescence imaging in Cy3 channel. The red arrow pointed to the target band. Uncropped gels were provided in Source Data. b The binding ability analysis of ZAJ4a, 7D12 and ZAJ4a-7D12 towards SACC-83 cells by CLSM in Cy3 channel. Merge: Cy3 channel and bright filed. Scale bar: 50 μm. c The binding ability analysis of ZAJ4a, 7D12 and ZAJ4a-7D12 by flow cytometry. d The last frame snapshot of EGFR/PTPRF/ZAJ4a-7D12 complex on membrane surface. EGFR, 7D12, PTPRF and ZAJ4a were colored in green, red, cyan and orange, respectively. The lipid bilayer was represented by black sticks. e Contact area of EGFR and 7D12, PTPRF and ZAJ4a in the system of EGFR/PTPRF/ZAJ4a-7D12 complex. f RMSD of each part of the complex. Both 7D12 and PTPRF were components of ZAJ4a-7D12. g Schematic illustration of ZAJ4a-7D12 induced artificial protein-proximity on the cell membrane. h Dual-color TIRF images of EGFR-mNeongreen (green) and PTPRF-Tdtomato (red). Scale bar: 5 μm. i Pearson’s correlation coefficients of EGFR-mNeongreen and PTPRF-Tdtomato. Data are obtained from Fig. 2h by using ImageJ software (n = 3 independent experiments). Data are shown as means ± SD, and p-values are determined by one-way ANOVA with Tukey’s multiple comparisons test in (i). The exact p-values are shown in the figure. Source data are provided as a Source Data file.

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