Western Blotting Analysis of Cell Signaling

Western blotting was performed as described earlier [45 (link)] using rabbit anti-RIG-I, rabbit anti-phospho-cdc2(Tyr15), rabbit anti-phospho-Chk2(Thr68) (Cell Signaling Technology Inc., Boston, MA, USA), and rabbit anti-DUSP6 (Abcam) monoclonal antibodies. In addition, rabbit anti-cyclinB1 and rabbit anti-CDK1(cdc2), and rabbit anti-Chk2 (Proteintech Group Inc., Wuhan, China) polyclonal antibodies were also used. All primary antibodies were diluted at 1:1000. The same membrane was stripped and reprobed with mouse anti-GAPDH, anti-β-actin, and anti-α-tubulin monoclonal antibodies (Proteintech Group Inc.) as internal controls.

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Li L., Lv L., Xu J.C., He Q., Chang N., Cui Y.Y., Tao Z.C., Zhu T, & Qian L.T. (2023). RIG-I Promotes Tumorigenesis and Confers Radioresistance of Esophageal Squamous Cell Carcinoma by Regulating DUSP6. International Journal of Molecular Sciences, 24(6), 5586.

Publication 2023

rabbit anti-RIG-I rabbit anti-DUSP6 mouse anti-GAPDH anti-β-actin

Actin Antibodies Cdc2 Dusp6 Gapdh Membrane Monoclonal antibodies Mouse Rabbit Rig i α tubulin

Corresponding Organization : University of Science and Technology of China

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Variable analysis

independent variables

Antibodies used for Western blotting: rabbit anti-RIG-I, rabbit anti-phospho-cdc2(Tyr15), rabbit anti-phospho-Chk2(Thr68), rabbit anti-DUSP6, rabbit anti-cyclinB1, rabbit anti-CDK1(cdc2), and rabbit anti-Chk2

dependent variables

Levels/expression of RIG-I, phospho-cdc2(Tyr15), phospho-Chk2(Thr68), DUSP6, cyclinB1, CDK1(cdc2), and Chk2 proteins

control variables

GAPDH, β-actin, and α-tubulin used as internal controls

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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