Investigating HDAC1, HSP90, NFKB2, IKKB, and TNF-α Expression

Western blotting and semi-quantitative analysis were performed as in our previous study [36 (link)]. To investigate the effect of GSZD on the expression levels of HDAC1, HSP90AA1, NFKB2, IKBKB and TNF-α proteins, HFLS were treated with 10 ng/mL of IL-1β in the presence of various concentrations of GSZD. Antibodies against the following proteins were used: HDAC1 (rabbit polyclonal antibody; dilution 1:1000; Abcam, Cambridge, UK), HSP90AA1 (rabbit polyclonal antibody; dilution 1:500; Abcam, Cambridge, UK), NFKB2 (rabbit monoclonal antibody; dilution 1:1000; Cell Signaling), IKKB (rabbit monoclonal antibody; dilution 1:1000; Abcam, Cambridge, UK), and TNF-α (rabbit polyclonal antibody; dilution 1:100; Abcam, Cambridge, UK). All experiments were performed in triplicate. The mean normalized protein expression ± SD was calculated from independent experiments.

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Guo Q., Mao X., Zhang Y., Meng S., Xi Y., Ding Y., Zhang X., Dai Y., Liu X., Wang C., Li Y, & Lin N. (2016). Guizhi-Shaoyao-Zhimu decoction attenuates rheumatoid arthritis partially by reversing inflammation-immune system imbalance. Journal of Translational Medicine, 14, 165.

Publication 2016

HDAC1 HSP90AA1 NFKB2 TNF-α

Antibodies Antibody Dilution Hsp90aa1 Il 10 Il 1β Monoclonal antibody Nfkb2 Protein Rabbit Tnf α

Corresponding Organization :

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing Jishuitan Hospital, Peking University, Wuhan University of Technology

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Variable analysis

independent variables

GSZD concentration

dependent variables

Expression levels of HDAC1, HSP90AA1, NFKB2, IKBKB and TNF-α proteins

control variables

Treatment with 10 ng/mL of IL-1β

controls

Positive control: Not specified
Negative control: Not specified

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