Antibody against inos

The four diterpenes, including ajudecumin A (1), Ajuforrestin B (2), (16S)-12,16-epoxy-11,14-dihydroxy-17(1 5 →16)-abeo-abieta-8,11,13-trien-7-one (3), and 14,15-dihydroajugapitin (4) were isolated from Ajuga ovalifolia var. calantha and identified by NMR (Chen et al. 2017b ), (Supplementary Figures S1 and S2). LPS from Escherichia coli 055:B5 and carrageenan were provided by Sigma (Shanghai, China). BAY 11-7082, CCK-8 agent, Griess agent was obtained from Beyotime (Haimen, China). Antibody against iNOS is the products of Cell Signalling Technology (Danvers, USA). Phospho- and total-ERK, phosphor- and total-p38 MAPK, phosphor- and total-JNK, phosphor- and total-IκBα antibodies were purchased from Signalway Antibody (Baltimore, USA). COX-2 and actin antibodies as well as HRP-conjugated secondary antibody were from Proteintech (Wuhan, China).

KRIBB11 was purchased from Calbiochem (San Diego, CA, USA). LPS was the product of Sigma (Saint Louis, MO, USA). Antibodies against IκB-α, NF-κB p65, p-NF-κB p65 (Ser536), Histone H2A, glyceral-dehyde-3-phosphate dehydrogenase (GAPDH), p-STAT1 (Tyr701), and STAT1 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibody against iNOS was purchased from Cell Signaling Technology or Abcam (Cambridge, MA, USA). Protein A/G PLUS-Agarose was the product of Santa Cruz Biotechnology (Santa Cruz, CA, USA). Other related agents were purchased from commercial suppliers. KRIBB11 was dissolved in dimethylsulfoxide (DMSO). The final concentration of DMSO was <0.05 %. All drugs were prepared as stock solutions, and stock solutions were stored at −20 °C.

Chemical standards for danshensu, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, salvianolic acid D rosmarinic acid, lithospermic acid, dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone IIA were purchased from Chengdu Must Bio-Tech Co. Ltd. (China). Ginsenosides Rg1, Rb1, Rh1, and Rd were purchased from Jilin University (China). Isolithospermic acid A, isolithospermic acid B and salvianolic acid G were isolated from Salvia miltiorrhiza in the authors’ laboratory. Their chemical structures (Supplementary Figure S1) were unambiguously identified by comparison of the NMR and MS spectra data with reported literatures. The purity of each reference compound was determined to be higher than 98% by normalization of the peak areas detected by HPLC-UV. LPS from Escherichia coli 055:B5 was purchased from Sigma (USA). Antibody against iNOS was obtained from Cell Signaling Technology (USA); antibodies against COX-2 and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, USA).