NF1 full-length or truncated proteins pulled down by KIPA beads were loaded to SDS-PAGE gel and separated by electrophoresis. The gel was then strained by Flamingo Fluorescent Gel Stain (Bio-Rad). Cell/tumor lysates were prepared and analyzed by Western blots as described previously (9 (link)). The antibodies target NF1 and GAPDH were from MilliporeSigma (MABN2557, Clone NF1-A376G3) and Santa Cruz Biotechnology, respectively. The antibodies against ERα (D8H8), ERK1/2, Phospho-ERK1/2 (T202/Y204), and β-tubulin (9F3) were all from Cell Signaling Technology.
Kim B.J., Zheng Z.Y., Lei J.T., Holt M.V., Chen A., Peng J., Fandino D., Singh P., Kennedy H., Dou Y., Chica-Parrado M.D., Bikorimana E., Ye D., Wang Y., Hanker A.B., Mohamed N., Hilsenbeck S.G., Lim B., Asirvatham J.R., Sreekumar A., Zhang B., Miles G., Anurag M., Ellis M.J, & Chang E.C. (2023). Proteogenomic Approaches for the Identification of NF1/Neurofibromin-depleted Estrogen Receptor–positive Breast Cancers for Targeted Treatment. Cancer Research Communications, 3(7), 1366-1377.
Other organizations :
Baylor College of Medicine, First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University, The University of Texas Southwestern Medical Center, Baylor Scott & White Health
Protein expression levels of NF1, GAPDH, ERα, ERK1/2, Phospho-ERK1/2, and β-tubulin in cell/tumor lysates
control variables
Experimental procedure for Western blot analysis as described previously (9 (link))
Antibodies used for Western blot analysis (NF1, GAPDH, ERα, ERK1/2, Phospho-ERK1/2, and β-tubulin)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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