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. 2021 Feb 3;11(2):200325. doi: 10.1098/rsob.200325

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© 2021 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 3. — Size exclusion chromatography of recombinant human γ-TuRC. (a) For SEC experiments, construct 1 (2xFLAG-GCP5, GCP6, GCP4, TUBG1 and ACTB) and construct 2 (MZT1, GCP2 and GCP3) were co-expressed with C-terminal Myc-His tagged γ-tubulin and purified via FLAG purification. (b) Chromatogram of a SEC run with ‘Superose 6 Increase (10/300) GL' column. Peak fractions (i (γ-TuRC peak), ii, iii) were analysed via immunoblotting (c,d). Red, brown and blue markers on x-axes (b) indicate borders of analysed fractions (i, ii, iii) in (d). Size markers thyroglobulin 669 kDa (V_t) and aldolase 158 kDa (V_a) are indicated. (c) FLAG elution left and ‘γ-TuRC peak' fraction (i, b) were probed by immunoblotting against the indicated antibodies. (d) Immunoblot analysis of ethanol precipitated peak fractions (i, ii, iii) from SEC experiment shown in (b). Equal amount of fraction volumes was loaded. See electronic supplementary material, figure S2 for uncropped images.