Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Endoplasmic Reticulum Chemical Chaperone 3‐Hydroxy‐2‐Naphthoic Acid Reduces Angiotensin II‐Induced Vascular Remodeling and Hypertension In Vivo and Protein Synthesis In Vitro

doi: 10.1161/JAHA.122.028201

Figure Lengend Snippet: Serum starved adventitial fibroblasts pretreated with 3‐hydroxy‐2‐naphthoic acid‐containing media or control vehicle‐containing media for 1 hour were treated with 1 μg/mL tunicamycin to induce endoplasmic reticulum stress and unfolded protein response. Immunoblotting was performed with the antibodies as indicated. A through E , Expression of unfolded protein response markers was examined and quantified. F and G , Phosphorylation of unfolded protein response markers was examined and quantified. Bar graph showed mean±SEM. 3HNA indicates 3‐hydroxy‐2‐naphthoic acid; ATF6, activated transcription factor 6; CHOP, C/EBP‐homologous protein; IRE1α, inositol requiring enzyme 1α; GRP78, 78‐kDa glucose‐regulated protein; eIF2α‐p, eukaryotic initiation factor 2α phosphorylated at Ser51; and IRE1α‐p, IRE1α phosphorylated at Ser724. Significance was indicated as * P <0.05, ** P <0.01, *** P <0.001, and **** P <0.0001.

Article Snippet: Primary antibodies used in this study were ATF6 (100 ng/mL, NBP1‐40256, Novus Biologicals), CHOP (C/EBP‐homologous protein) (1:10.000, 2895, Cell Signaling, Danvers, MA), GRP78 (70 ng/mL, 11587‐1‐AP, Proteintech, Rosemont, IL), IRE1α phosphorylated at Ser724 (200 ng/mL, MBS9610512, MyBioSource, San Diego, CA), IRE1α (40 ng/mL, sc‐390960, Santa Cruz Biotechnology, Dallas, TX), eIF2α phosphorylated at Ser51 (1:10.000, 3398, Cell Signaling), eIF2α (1:10.000, 2103, Cell Signaling), GAPDH (100 ng/mL, MAB374; Sigma‐Aldrich, St. Louis, MO), and β‐actin (6.1 ng/mL, 4970; Cell Signaling Technology).

Techniques: Western Blot, Expressing