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Figure 1
In vitro citrullination assays of different substrates. (a) ELISA assay for measuring citrullination activity of PAD4 and PAD6 on the histone H3 substrate. PAD6 (blue) and PAD4 (grey) were tested for histone H3 citrullination activity in the presence and absence of 1 µg ml−1 histone H3 (two independent experiments with n = 3). PAD4 citrullination activity was inhibited by 50 µM GSK484 inhibitor (green). All data were normalized to the citrullination signal of the substrate in the absence of PAD. Error bars indicate the standard deviation. (b) On-blot assay for measuring citrullination activity of PAD6 and PAD4 on different substrates. Citrullination of two substrates (histone H3 and cytokeratin 5) was evaluated in presence of PAD6 (blue, n = 7 for histone H3, n = 3 for cytokeratin 5) or PAD4 (grey, n = 5 for histone H3, n = 2 for cytokeratin 5) and normalized to the citrullination signal of the substrate in absence of PAD. PAD4 activity was inhibited by 50 µM GSK484 inhibitor (green, n = 4 for histone H3, n = 1 for cytokeratin 5).

IUCrJ
Volume 11| Part 3| May 2024| Pages 395-404
ISSN: 2052-2525
https://doi.org/10.1107/S2052252524002549