Supplementary Figure 6: Characterization of MYC and MYCS mutants in biological activity assays.
From: Multiple direct interactions of TBP with the MYC oncoprotein
a, Expression of MYC mutants (c-MYC, ΔMBII, MYCS, MYCSΔMBII) tagged with V5-epitope in TET21N cells, confirmed by Immunoblot analysis with V5 antibody, and β-actin antibody as loading control. b, Soft-agar assay, conducted for MYC mutants in cells expressing either Empty Vector (EV) or V5-tagged MYC or the MYCS or MYC without MBII (ΔMBII) or MYCSΔMBII. Average colony numbers for TET21N cells expressing either EV, MYC or the MYC mutants panel (normalized to MYC counts), seeded in soft-agar conditions and treated with 1µg/mL doxycycline-containing media prior to image collection and quantification. Error bars indicate SD from biological triplicate (n=3), *p< 0.05, One-way ANOVA test, Dunnett’s multiple comparisons correction. c, Sample images for the soft-agar analysis. d, Q-RT-PCR analysis of differentially regulated nucleolin (NCL) comparing different mutants to MYC. One-way ANOVA, *p<0.05, ***p<0.001 (n=3). e, Q-RT-PCR analysis of differentially regulated lactate dehydrogenase (LDHA) comparing different mutants to MYC. One-way ANOVA, *p<0.05, ***p<0.001 (n=3).
