Figure 1. TEAD1 forms foci in various RCC lines.

(A) Representative immunoblot showing TEAD1 and YAP1 protein levels in TEAD1 foci-positive versus TEAD1 foci-negative cell lines.

(B) Quantification of TEAD1 and YAP1 bands from (A), normalized to loading control. Data are presented as mean ± SD.

(C) Immunofluorescence imaging demonstrating TEAD1 foci formation in kidney tubule and renal carcinoma cell lines. Arrows indicate the TEAD1 foci. Scale bar = 5 μm. Nuclear boundaries are indicated by dotted lines.

(D) Quantification of TEAD1 foci in UOK121, UOK122, UOK342, and HK2 cell lines using 3D analysis. Data are presented as mean ± SEM.

(E-F) Maximum intensity projection from 3D images of TEAD1 (green) and Nucleolin (red) co-immunofluorescence in UOK342 and UOK121 (E), with quantification of TEAD1 foci nucleolar and perinuclear localization (n= 94–98 cells) (F).

(G) Full-bleach (top) Fluorescence Recovery after Photobleaching (FRAP) and half-bleach FRAP (bottom) of eGFP-TEAD1 UOK342 stable cell line over 10s. Scale bar = 2 μm

(H) Quantification of recovered fluorescence intensity of fully bleached TEAD1 foci (left, blue), half-bleached region (right, blue), and unbleached region (right, magenta). Shaded areas indicate the standard error of the mean (SEM) for each measurement (n = 10).

(I) Representative live-cell image of UOK342 TEAD1–HaloTag knock-in cells after treatment with DMSO control (top) or 1,6-hexanediol (1,6HD, bottom). HaloTag fluorescence was labeled with JF549 dye. Scale bar = 5 μm.

(J) Quantification of TEAD1 foci per cell in (I) (n = 45 for 1,6-hexanediol; n = 32 for DMSO). Data are shown as mean ± SEM; error bars represent the standard error of the mean. Statistical significance of all data was determined by one-way ANOVA: ns, p > 0.05; **, p ≤ 0.005; ****, p ≤ 0.0001.