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. 2023 Jun 19;16(6):dmm049132. doi: 10.1242/dmm.049132

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© 2023. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 4. — HCF1 processing by undifferentiated OGT^WT and OGT^C921Y in mESCs. (A) Immunoblot of HCF1 proteolytic fragments in the cytoplasm and nucleus of mESCs harbouring either OGT^WT or the OGT^C921Y variant. Lamin B1 was used as a marker for the nuclear fraction. This experiment was performed using three cell clones per genotype and repeated over two passages. (B) Quantification of HCF1 signal. HCF1 signals were normalised to those of β-actin. n=6 biological replicates. One-way ANOVA with Tukey’s comparison test; cytoplasmic fraction, P=0.99; nuclear fraction, P=0.54. (C) RT-PCR analysis of Hcf1 mRNA expression levels normalised to those of Gapdh, 18S (Rn18s) and Actb. Data points representing the mean expression calculated from three separate RT-PCR runs are shown. Each RT-PCR run was set up using several OGT^WT and OGT^C921Y lines as biological replicates. Unpaired two-tailed t-test, P=0.172. Error bars represent s.d. AU, arbitrary units. NS, not significant.