Phosphokinase Profiling and Immunofluorescence

Phosphokinase analysis was performed using the Proteome Profiler Human Phospho-Kinase Array Kit ARY003 (R&D Systems)^{53 (link)}. Array images were analyzed using the GeneTools image analysis software (Syngene). For Immunofluorescence, breast cancer cells (5 × 103) were seeded on chambered slides, allowed to grow for 24 h and treated as indicated and subjected to immunofluorescence analysis^{34 (link)}. Immunofluorescently stained cells were imaged using a Zeiss LSM510 Meta (Zeiss, Dublin, California, USA) laser scanning confocal system configured to a Zeiss Axioplan 2 upright microscope (Zeiss, Dublin, CA, USA).

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Nagalingam A., Siddharth S., Parida S., Muniraj N., Avtanski D., Kuppusamy P., Elsey J., Arbiser J.L., Győrffy B, & Sharma D. (2021). Hyperleptinemia in obese state renders luminal breast cancers refractory to tamoxifen by coordinating a crosstalk between Med1, miR205 and ErbB. NPJ Breast Cancer, 7, 105.

Publication 2021

Proteome Profiler Human Phospho-Kinase Array Kit ARY003 Zeiss LSM510 Meta Zeiss Axioplan 2 upright microscope

Breast cancer Cells Human Immunofluorescence Kinase Microscope Proteome

Corresponding Organization :

Other organizations : Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, University of Maryland, Baltimore

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Variable analysis

independent variables

Treatment conditions

dependent variables

Phosphokinase activity
Cellular localization and expression of proteins

control variables

Breast cancer cells (5 × 10^3) seeded on chambered slides
Cells allowed to grow for 24 h

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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