Journal: Frontiers in Endocrinology

Article Title: Biochanin A Alleviates Cerebral Ischemia/Reperfusion Injury by Suppressing Endoplasmic Reticulum Stress-Induced Apoptosis and p38MAPK Signaling Pathway In Vivo and In Vitro

doi: 10.3389/fendo.2021.646720

Figure Lengend Snippet: Biochanin A ameliorated endoplasmic reticulum stress and inhibited the phosphorylation activity of p38 in OGD/R neurons. (A) Protein expression of GRP78 in primary cortical neurons with OGD/R induction. β-actin were used as internal controls. (B) Protein expression of CHOP in primary cortical neurons with OGD/R induction. PCNA were used as internal controls. (C) Effects of biochanin A on the phosphorylation activity of p38 in primary cortical neurons after OGD/R. The ratio of p-p38 to total p38 was used to evaluate protein activity. Results were presented as mean ± SE, n = 3. # P < 0.05, the OGD/R group vs. the control group; *P < 0.05, vs. the OGD/R group.

Article Snippet: These blots were then blocked for 1 h using 5% non-fat milk in TBST (tris-buffered saline containing 0.1% Tween-20) at 37°C, after which they were incubated at 4°C overnight with appropriate primary antibodies: anti-GRP78 (sc-13968, Santa Cruz Biotechnology, Dallas, TX, USA, 1:200), anti-CHOP (15204-1-AP, Proteintch Group, Wuhan, China, 1:200), anti-p38 (9212, Cell Signaling Technology, Danvers, USA, 1:1000), anti-p-p38 (4511, Cell Signaling Technology, Danvers, USA, 1:1000), anti-caspase-3 (19677-1-AP, Proteintch Group, Wuhan, China, 1:500), anti-caspase-12 (ab62484, Abcam, Cambridge, UK, 1:500), anti-Bcl-2 (12789-1-AP, Proteintch Group, Wuhan, China,1:1000), anti-Bax (50599-2-lg, Proteintch Group, Wuhan, China, 1:2000), anti-β-actin (TA-09, ZSGB-Bio, Beijing, China, 1:1000), anti-α-Tubnin (AF7010, Abcam, Cambridge, UK, 1:1000), anti-PCNA (A01040, Abbkine, Wuhan, China, 1:1000).

Techniques: Activity Assay, Expressing