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. 2020 Dec 21;118(1):e2007526118. doi: 10.1073/pnas.2007526118

Fig. 5.

Fig. 5.

Replacing the p14/p22 branched actin nucleator with a formin is sufficient to drive cell-cell fusion. (A) Diagram of p14/p22 chimera with FKBP at C terminus and P149A mutation with FRB-tagged ΔGBD-mDia2. (B) Confocal images of p14/p22 chimera P149A FKBP-mCherry (magenta) coexpressed with FRB-ΔGBD-mDia2 and Lifeact-GFP (green) cell at 0 min and 10 min after addition of 500 nM rapalog. Filopodia-like protrusions before and after rapalog addition is denoted with arrows. Box regions magnified. (C) Representative confocal merged image of a filopodia-like protrusion from cell expressing p14/p22 chimera-P149A-FKBP-mCherry (magenta), Lifeact-GFP (green), and FRB-ΔGBD-mDia2 10 min after addition of 500 nM rapalog. Each channel is shown with p14/p22 chimera-P149A-FKBP-mCherry (fire) and Lifeact-GFP (green). Average normalized fluorescence intensity of p14/p22 chimera-P149A-FKBP-mCherry along the length before (n = 33 filopodia-like protrusions) and 10 min after (n = 36 filopodia-like protrusions) addition of 500 nM rapalog. SD above and below the average are shown. ****P < 0.0001 by Student’s t test on the last datapoint. (D) Distribution of number of nuclei in p14/p22 chimera-P149A-FKBP– and FRB-ΔGBD-mDia2–expressing cells with and without rapalog from three independent transfections, with mean number of each replicate, average from three independent transfections shown. P values are two-tailed two-sample Student’s t test where ***P < 0.001.