Lnc-17-92 mediates the assembly of a MYC-WDR82 transcriptional complex, leading to transcriptional and epigenetic activation of ACACA. (A) Schematic of integrated BioID and coimmunoprecipitation assay followed by mass-spectrometry analysis (Co-IP/MS) assays to explore the MYC-protein interacting network in the presence or absence of lnc-17-92 depletion. (B) Western blot analysis of WDR82 in RPPD material precipitated with lnc-17-92^TV1 or lnc-17-92^TV2 or with control RNA; 5% input is used as a reference. (C) RNA Y3H using WDR82 as hybrid protein 2 and, as hybrid RNAs, either a negative control RNA (−) or lnc-17-92^TV1 or lnc-17-92^TV2. Red arrows indicate yeast colony growth. (D) ChIP-qPCR analysis of H3K4me3 occupancy at ACACA promoter after silencing of WDR82 with a siRNA pool (n-4) in H929 (24-hour time point). Data are represented as % of input chromatin. (E) ChIP-qPCR analysis of MYC occupancy at the ACACA promoter after silencing of WDR82 with a siRNA pool (n-4) (24-hour time point). Data are represented as % of input chromatin. (F) qRT-PCR analysis of ACACA mRNA after silencing of WDR82 with a siRNA pool (n-4) (48-hour time point). Raw Ct values were normalized to GAPDH mRNA and expressed as ΔΔCt values calculated using the comparative cross threshold method. ACACA expression levels in cells transfected with NC were set as an internal reference. (G) ChIP-qPCR analysis of WDR82-GFP occupancy at the ACACA promoter in AMO1 exposed for 24 hours to gymnotic ASO1. Data are represented as % of input chromatin. Western blot analysis of WDR82-GFP from paired samples. α-Tubulin was used as the protein loading control. (H) ChIP-qPCR analysis of H3K4me3 occupancy at the ACACA promoter in AMO1 and H929 exposed for 24 hours to gymnotic ASO1. Data are represented as % of input chromatin. (I) Western blot analysis of H3, H3H3K4me1, H3H3K4me2, and H3H3K4me3 in AMO1 and H929 exposed for 24 hours to gymnotic ASO1. Lamin A/C was used as the protein loading controls (nuclear lysates). ∗P < .05, Student t test.