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. 2021 May 13;184(10):2649–2664.e18. doi: 10.1016/j.cell.2021.03.031

Figure S2.

Figure S2

RAS adaptor protein GRB2 is recruited to EML4-ALK cytoplasmic protein granules, related to Figure 2

(A) Live-cell confocal imaging of mTagBFP2::EGFR L858R (oncogenic EGFR) expressed in human epithelial cell lines (Beas2B) with endogenous mNeonGreen2-tagging of GRB2, GAB1, and SOS1. Arrows denote plasma membrane enrichment of GRB2, GAB1, and SOS1. Representative images from at least 30 cells analyzed per condition in 3 independent experiments.

(B) Live-cell confocal imaging of mTagBFP2::EML4-ALK and mEGFP::GRB2 upon dual expression in Beas2B cells. White arrows indicate a representative EML4-ALK cytoplasmic protein granule with local enrichment of GRB2 (multiple non-highlighted granules also show colocalization between EML4-ALK and GRB2). Images are representative of 100 analyzed cells in total over 3 independent experiments.

(C) FRAP experiments performed in human epithelial cells (Beas2B) with endogenous mNeonGreen2-tagging of GRB2, upon expression of mTagBFP2::EML4-ALK. Graph displays individual recovery of fluorescence intensity after photobleaching of GRB2 enriched at EML4-ALK granules, t denotes time in seconds, intensity in arbitrary units (a.u.) normalized to 1. N = 30 cells.