Western Blot Antibody Validation

Western blotting was performed as previously described 28 (link). The primary antibodies for PD-L1 (CST #13684), IRF1 (CST #8478), STAT1 (CST #14994), p-STAT1 (CST #9167), GZMB (CST #17215), β-actin (CST #4970) and the matching horseradish peroxidase (HRP) conjugated secondary antibodies were purchased from Cell Signaling Technologies (CST). Immobilon Western Chemiluminescent HRP Substrate (Millipore, Billerica) was added, and the bands were imaged with the Chemidoc XRS+ electrophoretic imaging system (Bio-Rad). Density scanning of each protein band was performed using Image Lab software (Bio-Rad). The results were normalized to β-actin.

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Li R., Zhan Y., Ding X., Cui J., Han Y., Zhang J., Zhang J., Li W., Wang L, & Jiang J. (2024). Cancer Differentiation Inducer Chlorogenic Acid Suppresses PD-L1 Expression and Boosts Antitumor Immunity of PD-1 Antibody. International Journal of Biological Sciences, 20(1), 61-77.

Publication 2024

PD-L1 STAT1 p-STAT1 β-actin Immobilon Western Chemiluminescent HRP Substrate Chemidoc XRS+ electrophoretic imaging system Image Lab software

Corresponding Organization :

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Peking Union Medical College Hospital, Kunming Institute of Botany, Chinese Academy of Sciences, Science and Technology Department of Sichuan Province, Capital Medical University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of PD-L1, IRF1, STAT1, p-STAT1, GZMB

control variables

β-actin for normalization of protein expression

controls

No positive or negative controls specified

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