Fig. 5: ARMC9 colocalizes with TOGARAM1 and CSPP1 and enhances their effects on MT dynamics.
a–c, Fields of view (left; scale bar: 2 µm) and kymographs (right; scale bars: 2 µm and 60 s) illustrating MT dynamics from GMPCPP-stabilized seeds with 20 nM GFP–EB3 or mCherry–EB3 in indicated conditions. Assays were repeated independently three times. Blue arrowheads, ARMC9 and TOGARAM1 colocalization. d, Parameters of MT plus-end dynamics in the presence of 20 nM EB3 in combination with indicated concentrations of TOGARAM1 constructs and ARMC9 (from kymographs shown in a,b and Figs. 1c,p and 4e). For growth rates, bars represent the pooled data from three independent experiments. Total number of growth events: EB3 alone, n = 938; EB3 with TOGARAM1, n = 861; EB3 with TOGARAM1 and ARMC9, n = 342; EB3 with TOGARAM1 123′4′, n = 337; EB3 with TOGARAM1 123′4′ and ARMC9, n = 325. For transition frequencies, bars represent averaged means from three independent experiments. Error bars represent the s.e.m. ****P < 0.0001 (Kruskal–Wallis test followed by Dunn’s post hoc test). e, Fields of view (left; scale bar: 2 µm) and kymographs (right; scale bars: 2 µm and 60 s) illustrating MT dynamics from GMPCPP-stabilized seeds with 20 nM GFP–EB3 and CTM proteins indicated. Yellow arrowheads, colocalization. f, Parameters of MT plus-end dynamics with 20 nM EB3 in combination with indicated concentrations of ciliary top module proteins (from kymographs shown in e and Fig. 1c,n). For growth rate and pause duration, bars represent the pooled data from three independent experiments. Total number of growth events, pauses: EB3 alone, n = 938, 0; EB3 with CSPP1, n = 1,715, 273; EB3 with CSPP1 and ARMC9, n = 627, 315. For transition frequencies and dynamic state, bars represent the averaged means from three independent experiments. Error bars represent the s.e.m. ****P < 0.0001 (Kruskal–Wallis test followed by Dunn’s post hoc test). g,h, Fields of view (left; scale bar: 2 µm) and kymographs (right; scale bars: 2 µm and 60 s) illustrating MT dynamics from GMPCPP-stabilized seeds with 20 nM GFP–EB3 in indicated conditions. Assays were repeated independently three times. Yellow arrowheads, colocalization. i, Schematic model showing TOGARAM1, ARMC9 and CSPP1 interaction domains.
