Figure 2.

NF2 mutant showed restricted binding to the plasma membrane

(A) pEGFP-NF2-WT, pEGFP-NF2-mut and pEGFP were expressed in HEK293T cells, respectively. The immunofluorescence pattern of transfected HEK293T cells was double-labeled with CellMask™ and GFP. Scale bar, 50μm. See also Figure S2A.

(B) The Lipid dot-blot assay confirmed the interaction between PA with NF2. See also Figure S2B.

(C) Time-series fluorescence microscopy analysis of GFP-NF2 puncta. Bottom row shows zoom-in view of two fusing puncta. Scale bar, 10μm (top) and 1μm (bottom). See also Video S1.

(D) Representative micrographs of GFP-NF2 puncta before and after photobleaching. Scale bar, 0.2μm. See also Figure S3.

(D′) Quantification of fluorescence intensity recovery in the bleached region of GFP-NF2 puncta. Error bars, SEM of three independent experiments.

(E) NF2-mut contained an IDR.

(F) GFP-NF2-mutΔIDR were expressed in HEK293T cells. The immunofluorescence pattern of NF2-mutΔIDR puncta. Scale bar, 20μm.

(F′) Quantification of puncta per cell. Error bars, SEM of three micrographs. ^nsP>0.05, Student’s t test.

(G) Time-series fluorescence microscopy analysis of NF2-mutΔIDR puncta. Bottom row shows zoom-in view of two fusing puncta. Scale bar, 10μm (top) and 1μm (bottom). See also Video S2.

(H) Representative micrographs of NF2-mutΔIDR puncta before and after photobleaching. Scale bar, 0.2μm.

(H′) Quantification of fluorescence intensity recovery in the bleached region of NF2-mutΔIDR puncta. Error bars, SEM of three independent experiments.

(I) No in vitro liquid droplets of either NF2-WT, NF2-mut or NF2-mutΔIDR proteins.