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. 2019 Apr 3;316(6):F1180–F1190. doi: 10.1152/ajprenal.00089.2019

Fig. 1.

Fig. 1.

A: recombinant human peptidyl arginine deiminase-4 (PAD4) preferentially citrullinates IKKγ-glutathione S-transferase (GST) [NF-κB essential modulator (NEMO)]. Recombinant human IKKα-GST, IKKβ-GST, or IKKγ-GST (0.5 μg) was incubated with vehicle (saline) or with recombinant human PAD4 (0.5 μg) for 3 h and probed for citrullinated NEMO and total NEMO (representative images from 4 experiments). Note that IKKα-GST and IKKβ-GST showed larger bands at ~100 kDa, whereas IKKγ-GST was smaller with a molecular mass of ~75 kDa. B and C: recombinant human PAD4 citrullinates NEMO in human proximal tubule cells (HK-2; B) and mouse proximal tubule cells (C). HK-2 cells or freshly isolated mouse proximal tubule cells were treated with vehicle or with recombinant human PAD4 (10 μg/ml) for 3 h and probed for citrullinated NEMO and total NEMO. Bar graphs show band intensity quantifications demonstrating increased NEMO citrullination by recombinant human PAD4 in both experimental conditions. *P < 0.05 vs. vehicle (n = 4–6 experiments). Error bars represent 1 SE.