Western Blot Analysis of Protein Markers

Whole cell lysates were prepared as previously described.18 (link) Equal amounts of protein were boiled, separated on 10% SDS-PAGE, transferred onto a PVDF membrane, and visualized using an enhanced chemiluminescence kit (Millipore, MA, USA). Antibodies against PTPRO (ab150834), STAT3 (Y705) (ab76315), STAT3 (S727) (ab30647), STAT3 (ab68153), STAT2 (Tyr690) (#88410; Cell Signaling), STAT2 (ab134192), STAT1 (Y701) (ab30645), STAT1 (ab31369), JAK2 (Y1007+Y1008) (ab32101), JAK2 (ab108596), c-MYC (s62) (ab51156), c-MYC (ab39688), human PD-L1 (ab205921), mouse PD-L1 (ab233482), and β-actin (ab6276) were purchased from Abcam. All uncropped western-blot figures are presented in online supplementary figures s13‒16.

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Zhang W., Liu Y., Yan Z., Yang H., Sun W., Yao Y., Chen Y, & Jiang R. (2020). IL-6 promotes PD-L1 expression in monocytes and macrophages by decreasing protein tyrosine phosphatase receptor type O expression in human hepatocellular carcinoma. Journal for Immunotherapy of Cancer, 8(1), e000285.

Publication 2020

enhanced chemiluminescence kit ab76315 ab30645 ab32101 ab108596 (ab51156 ab39688 ab205921 ab233482 ab6276

Actin Antibodies C myc Cell Chemiluminescence Human Jak2 Membrane Mouse Pd l1 Protein Ptpro Pvdf Sds page Stat1 Stat2 Stat3 Western blot

Corresponding Organization : Jiangsu Cancer Hospital

Other organizations : Jiangsu University, First People's Hospital of Kunshan

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Variable analysis

independent variables

Whole cell lysates preparation method

dependent variables

Protein levels of PTPRO, STAT3 (Y705), STAT3 (S727), STAT3, STAT2 (Tyr690), STAT2, STAT1 (Y701), STAT1, JAK2 (Y1007+Y1008), JAK2, c-MYC (s62), c-MYC, human PD-L1, mouse PD-L1, and β-actin

control variables

Equal amounts of protein were used
Proteins were boiled and separated on 10% SDS-PAGE
Proteins were transferred onto a PVDF membrane
Proteins were visualized using an enhanced chemiluminescence kit

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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