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. 2013 Dec 26;8(12):e84462. doi: 10.1371/journal.pone.0084462

Figure 1. Phosphorylation of ERα at serine 216.

Figure 1

(A) Map to show localization of serine 216 in ERα and threonine 38 in CAR. (B) In vitro phosphorylation of serine 216. Purified glutathione S transferase (GST)-mERα wild type (WT) and GST-S216A were incubated with or without protein kinase C (PKC) and Western blots were performed using an anti-P-Ser-216 (αP-S216) or anti-GST antibody. (C) Western blot analysis of whole extracts (20 μg protein/well) prepared from the mouse uterus at the estrus stage with αP-S216 or an anti-ERα antibody (αERαN or αERαC). (D) Uterine sections were immune-stained using αP-S216 antibody or normal IgG. (E) Uterine sections to determine staining specificity of αP-S216. αP-S216 antibody was mixed with phospho- or non-phospho-peptides prior to incubation with sections.