EML4-ALK membraneless cytoplasmic protein granules recruit RAS-activating complex GRB2/SOS1/GAB1 in situ
(A) Live-cell imaging of Beas2B cells with endogenous mNG2-tagging of signaling proteins ± mTagBFP2::EML4-ALK.
(B) Signaling protein colocalization with EML4-ALK granules. 100 total cells per condition n = 3.
(C) Fold-enrichment of signaling proteins at EML4-ALK granules ±24 h treatment with 1 μM crizotinib (criz). ∗∗p < 0.01, paired t test. Orange line denotes zero enrichment.
(D) Anti-ALK IF images in H3122 cells with endogenous mNG2-tagging of GRB2 representative of 20+ cells, n = 3. Scale bar, 5 μM.
For (A) and (D), arrows indicate a representative EML4-ALK granule with local enrichment of signaling proteins (multiple non-highlighted granules also show colocalization). For all panels, error bars represent ±SEM.
See also Figure S2.