Total cell lysates from human RMS cells and xenografts were obtained following lysis in SDS lysis buffer. Western blot analysis was performed similar to Ignatius et. al., 2017 (21 (link)). Membranes were developed using an ECL reagent (Western Lightning Plus ECL, PerkinElmer; or sensitive SuperSignal West Femto Maximum Sensitivity Substrate, Thermo Scientific). Membranes were stripped, rinsed, and re-probed with the respective internal control antibodies. List of primary and secondary antibodies is included in supplementary data (Supplemental Table 2 ).
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Western Blot Analysis of RMS Cells
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Corresponding Organization : The University of Texas Health Science Center at Houston
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Variable analysis
independent variables
- Type of cells (human RMS cells and xenografts)
- Protein expression levels (as measured by Western blot analysis)
- Lysis buffer (SDS lysis buffer)
- Western blot protocol (similar to Ignatius et al., 2017)
- Detection method (ECL reagent, Western Lightning Plus ECL or SuperSignal West Femto Maximum Sensitivity Substrate)
- Internal control antibodies (as listed in supplementary data)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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