Figure 2

In vitro G4 folding assessment and CNBP binding of the Hs791 and Hs2160 PQSs. (A) DNA intrinsic fluorescence (in arbitrary units (a.u.), left panels) and CD (right panels) spectra of synthetic oligonucleotides representing the PQSs Hs791 (top) and Hs2160 (bottom). The potassium ions concentrations are indicated in the insets. (B) Thermal denaturation curves of synthetic oligonucleotides representing Hs791 (black) and Hs2160 (blue) obtained by monitoring the ellipticity at 260 nm as a function of temperature. Dots correspond to experimental data; lines are fits to a sigmoid function. Estimated melting temperatures (Tm) are informed in the plot. (C) Table reporting K_d ± SEM values obtained in EMSAs for each probe in both folding states. In all cases 3 independent repeats were performed for each probe and folding state. (D) ChIP assays performed on CNBP-eGFP expressing HeLa cells using anti-GFP or anti-IgG control antibodies. Bars represent the average enrichment of qPCR-amplified Hs2160 and Hs791 sequences in immunoprecipitated chromatin with anti-GFP antibody or with control antibody. GAPDH was used as amplification control (gene not regulated by CNBP). In all cases, bars represent the mean ± SD of three independent biological replicates. ***P < 0.001, NS not significant, t test.