RPPA Analysis of Myc-tagged KPC1 in Melanoma

Protein lysate from melanoma lines (IM-0223 and MH-0331) stably transfected with either Myc-tagged KPC1 or empty-vector (OriGene) was extracted as previously described (32 (link)), and RPPA analysis was performed by the CCSG-supported RPPA Core Facility at the University of Texas MD Anderson Cancer Center (32 (link)). A total of 232 authenticated Abs for total protein expression and 62 Abs for protein phosphorylation were analyzed in this study. A list of the Abs can be accessed from https://www.mdanderson.org/education-and-research/resources-for-professionals/scientific-resources/core-facilities-and-services/functional-proteomics-rppa-core/antibody-lists-protocols/functional-proteomics-reverse-phase-protein-array-core-facility-antibody-lists-and-protocols.html (Ab set 107). Heat maps and Volcano plots were generated from load-corrected log2 data using MultiExperiment Viewer (MeV) v4.7.1. Differences in protein expression between groups were determined using student’s t test with a two-sided p-value < 0.05.

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Iida Y., Ciechanover A., Marzese D.M., Hata K., Bustos M., Ono S., Wang J., Salomon M.P., Tran K., Lam S., Hsu S., Nelson N., Kravtsova-Ivantsiv Y., Mills G.B., Davies M.A, & Hoon D.S. (2017). Epigenetic Regulation of KPC1 Ubiquitin Ligase Effects the NF-κB Pathway in Melanoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(16), 4831-4842.

Publication 2017

empty-vector

Antibody Cancer Maps Melanoma Phosphorylation Protein Protein array Student Vector

Corresponding Organization : Saint John's Health Center

Other organizations : Technion – Israel Institute of Technology, The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

Myc-tagged KPC1 transfection
Empty-vector (OriGene) transfection

dependent variables

Total protein expression
Protein phosphorylation

control variables

Protein lysate from melanoma lines (IM-0223 and MH-0331)

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