Fig. 2: Direct binding of Ndc80C to CENP-T facilitates the recruitment of Mis12C into the kinetochore.
From: Recruitment of two Ndc80 complexes via the CENP-T pathway is sufficient for kinetochore functions
a Amino acids alignment of CENP-T between various species. Potential phosphorylation sites (T72 and S88) are marked. b Nuf2-GFP levels at kinetochores in cKO-CENP-T cells expressing mScarlet-CENP-C∆73 and either CENP-TWT, CENP-TΔ90, or CENP-TT72A-S88A in the presence of Tet for 30 h. None indicates parental cKO-CENP-T/CENP-C∆73/Nuf2-GFP cells. Subtract values are presented using signal intensities of None. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated by one-way ANOVA followed by Tukey’s test. c Amino acids alignment of CENP-T between various species. Mutated S/T residues are marked. d GFP-Dsn1 levels at kinetochores in cKO-CENP-T cells expressing mScarlet-CENP-C∆73 and CENP-T8A in the presence of Tet for 48 h. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated by two-tailed Welch’s t-test. e The growth of cKO-CENP-T cells expressing mScarlet-CENP-C∆73 and either CENP-T8A or CENP-TWT in the absence or presence of Tet. f GFP-Dsn1 levels at kinetochores in cKO-CENP-T cells expressing mScarlet-CENP-C∆73 and either CENP-TWT, CENP-TΔ121-240, or CENP-TΔ90 in the presence of Tet for 30 h. None indicates parental cKO-CENP-T/CENP-C∆73/GFP-Dsn1 cells. Subtract values with None are presented. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated as in b. g GFP-Dsn1 levels at kinetochores in cKO-Spc25 cells expressing mScarlet-CENP-C∆73 in the absence or presence of Tet for 18 h. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated as in d. h GFP-Dsn1 levels at kinetochores in CENP-C KO cells expressing HA-CENP-C∆73 (CENP-C∆73 cells) in the presence of nocodazole or MG132. CENP-T was immuno-stained. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated as in d. i Nuf2-GFP levels at kinetochores in CENP-C∆73 cells in the presence of nocodazole or MG132. CENP-T was immuno-stained. DNA was stained with DAPI. Error bars indicate the mean and standard deviation. p values were calculated as in d.
