Fig. 2.

Effect of Vit D on PAD1–3 protein levels and PAD activity in NHKs. NHKs were treated with Vit D (with or without 1.5 mM calcium) or DMSO as the vehicle, as mentioned on the top of the panels, and harvested at the indicated time. a,c Proteins of the “TE-NP40” (a) and “Laemmli” (c) extracts were separated by SDS-PAGE, transferred to nitrocellulose membranes and stained with Ponceau S. b Immunodetection of PAD1–3 and actin in “TE-NP40” extracts. d Immunodetection of deiminated proteins using the anti-citrulline AMC antibody in the “TE-NP40” (top) and “Laemmli” (bottom) extracts. Molecular masses are reported in kDa on the right. Positive controls (Co) correspond either to purified recombinant human PAD1, PAD2 and PAD3, and to human epidermal extracts (b) or to deiminated fibrinogen (d)