Journal: The Journal of Biological Chemistry

Article Title: The 14-3-3γ isoform binds to and regulates the localization of endoplasmic reticulum (ER) membrane protein TMCC3 for the reticular network of the ER

doi: 10.1016/j.jbc.2022.102813

Figure Lengend Snippet: Phosphorylated serine 15 in the deduced 14-3-3 binding motif is required for potent binding to 14-3-3γ . A , the N-terminal region of TMCC3 highlighting the deduced 14-3-3 binding motifs. Serine residues being the possible phosphorylation sites in three deduced 14-3-3 binding motifs are indicated in red . Arginine, serine, and proline residues at −3, 0, and +2 positions, respectively, in the deduced 14-3-3 binding motif predicted with the highest score are underlined. B , serine 15 is required for potent binding to 14-3-3γ. Indicated combinations of HA-TMCC3-D1, HA-TMCC3-D1-S15A, HA-TMCC3-D1-S15/25/46A, and Flag-14-3-3γ were transfected into HEK293 cells, followed by immunoprecipitation with the anti-Flag mAb. The samples were immunoblotted with the anti-HA mAb and the anti-Flag pAb. The arrowheads indicate the bands of interest. The asterisk s indicate the nonspecific bands from the light chain of IgG. C , serine 15 is phosphorylated. HA-TMCC3-D1, HA-TMCC3-D1-S15A, and HA-TMCC3-D1-S15/25/46A were transfected into HEK293 cells, followed by immunoprecipitation with the anti-HA mAb. The samples were immunoblotted with the anti-phospho-14-3-3 binding motif pAb and the anti-HA pAb. The asterisk indicates the nonspecific bands from the light chain of IgG. TMCC3, transmembrane and coiled-coil domain family 3.

Article Snippet: The antibodies were purchased from the following commercial sources: mouse anti-HA mAb (BioLegend; Cat. No. 901514), rabbit anti-HA pAb (Sigma-Aldrich; Cat. No. H6908), rat anti-HA mAb (Roche; Cat. No. 11867423001), mouse anti-FLAG mAb (Sigma-Aldrich; Cat. No. F1804), rabbit anti-FLAG pAb (Sigma-Aldrich; Cat. No. F7425), rabbit anti-TMCC3 pAb (Sigma-Aldrich; Cat. No. HPA014272), rabbit anti-CLIMP-63 pAb (Bethyl Laboratories; Cat. No. A302–257A), mouse anti-α-tubulin mAb (Sigma-Aldrich; Cat. No. T6199), mouse anti-14-3-3 mAb (Santa Cruz Biotechnology; Cat. No. sc-1657), rabbit anti-phospho 14-3-3 binding motif pAb (Cell Signaling; Cat. No. 9601), mouse anti-PDI mAb (Abcam; Cat. No. ab2792), rat anti-GFP mAb (Nacalai; Cat. No. 04404–84), rabbit anti-GFP pAb (MBL; Cat. No. 598), and mouse anti-GAPDH mAb-HRP-DirecT (MBL; Cat. No. M171–7).

Techniques: Binding Assay, Transfection, Immunoprecipitation

Journal: Molecular Endocrinology

Article Title: Interferon Tau Regulates PGF2α Release from the Ovine Endometrial Epithelial Cells via Activation of Novel JAK/EGFR/ERK/EGR-1 Pathways

doi: 10.1210/me.2010-0205

Figure Lengend Snippet: List of antibodies used

Article Snippet: 26 , p-Serine , Rabbit polyclonal , Ser , 1:1000 , 9601 , Cell Signaling.

Techniques: Concentration Assay