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. Author manuscript; available in PMC: 2022 Feb 28.
Published in final edited form as: Oncogene. 2021 Oct 30;41(2):220–232. doi: 10.1038/s41388-021-02086-9

Fig. 3. Proximity-based labeling by SNX3-TurboID.

Fig. 3

A Western blotting analysis on lysates from cells transfected with 2XHA-TurboID or 2XHA-TurboID-SNX3 constructs for proximity-based labeling assay. For HeLa cells, immune-precipitated (IP) samples were immunoblotted with EGFR and HA antibodies. In parallel, the 2XHA-TurboID or 2XHA-TurboID-SNX3 and EGFR-GFP constructs were tranfected to HEK293 cells. IP samples were immunoblotted with HA and GFP antibodies. GFP-antibody was used to detect the EGFR-GFP in the IP samples. HA-antibody was used to verify the expression of TurboID or TurboID-SNX3 fusion protein for both cells. Whole-cell lysates (WCL) were used to show the equal loading of proteins. Blots are representative of 3 independent experiments. B Western blotting analysis on lysates from HeLa cells transfected with 2XHA-TurboID or 2XHA-TurboID-SNX3 constructs and treated with EGF as indicated. Biotinylated EGFR levels were detected. HA-antibody was used to verify the expression of TurboID or TurboID-SNX3 fusion protein. WCL shows EGFR, phosphotyrosine (pY), and ACTIN levels. Blots and the graph for densitometry analysis represent 3 independent experiments. One-way ANOVA and Dunnett’s multiple comparison tests were used to measure statistical significance, ***p < 0.001.