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. 2022 Oct 4;25(11):105275. doi: 10.1016/j.isci.2022.105275

Figure 3.

Figure 3

NF2 mutant restricted the recruitment of LATS to the plasma membrane and formed LLPS of NF2-LATS in the cytoplasm

(A) The pRFP-LATS only, pRFP-LATS and pEGFP-NF2-WT, pRFP-LATS and pEGFP-NF2-mut, pRFP-LATS and pEGFP-NF2-mut BDdel were expressed in HEK293T cells, respectively. The immunofluorescence pattern of in transfected HEK293T cells. The number (A′) and size (A″) of puncta were quantified. Scale bar, 20μm. See also Figure S4.

(B) Phase-separation assays of LATS, LATS and NF2-WT, LATS and NF2-mut proteins in vitro. Scale bar, 20μm.

(C) Time-series fluorescence microscopy analysis of LATS, LATS and NF2-WT, LATS and NF2-mut. Scale bar, 20μm.

(D, D′) in vitro FRAP assay and quantification of LATS and NF2-WT puncta.

(E, E′) in vitro FRAP assay and quantification of LATS and NF2-mut puncta.

(F, F′) in vitro FRAP assay and quantification of LATS puncta. Time 0s indicates the photobleaching pulse. Scale bar, 20μm. Error bars, SEM of three independent experiments. Data are represented as mean ± SEM ∗∗∗p < 0.001, Student’s t test.