TPD54 homo- and heteromerizes using its coiled-coil domain. (A) Schematic representation of TPD54 and a graph showing the coiled-coil probability for WT and L53P,L67P (PP) mutant. Pink, coiled-coil domain; light blue, conserved region. For coiled-coil domain prediction, the amino acid sequences of TPD54 isoform 1 WT or TPD54(L53P,L67P) were analyzed by PCOILS with window size of 28 (Gruber et al., 2006). (B) Western blot (WB) showing immunoprecipitation (IP) of FLAG-TPDs and coimmunoprecipitation of GFP-FKBP, GFP-FKBP-TPD54 (WT) or GFP-FKBP-TPD54(L53P,L67P; PP). (C) Representative confocal image of mCherry-FKBP-TPD54 (TPD54 WT) or mCherry-FKBP-TPD54(L53P,L67P; TPD54 PP) expressed in HeLa cells. Scale bar, 10 µm. (D) Scatter dot plot to show the spatiotemporal variance (STV) of the indicated constructs. Dots, individual cells; black bars, mean ± SD. The mean ± SD for mCherry-FKBP (control) is also shown as a black line and gray zone. Values for R159E are shown as a negative control. WT and PP, P = 1.9 × 10⁻⁶; Dunnett’s post hoc test using mCherry-FKBP as control.