Journal: Cell reports

Article Title: linc-ADAIN , a human adipose lincRNA, regulates adipogenesis by modulating KLF5 and IL-8 mRNA stability

doi: 10.1016/j.celrep.2024.114240

Figure Lengend Snippet: (A) Regulatory features at the linc-ADAIN locus, RNA-seq coverage (human adipocytes), transcription factor binding, and active histone modification markers. (B) Tissue expression of linc-ADAIN from GTEx (gene mean transcripts per million [TPM]). (C and D) Induction of linc-ADAIN by the PPARγ agonist rosiglitazone (10 μM) (C) and during adipocyte differentiation in vitro (D) ( N = 3 in triplicate). (E) Cellular fractionation of primary ASC adipocytes. qPCR of MALAT1, GAPDH, and linc-ADAIN of nuclear and cytoplasmic fractions. Data were normalized by averaging of loading controls GAPDH, β-ACTIN, MALAT1, U6, and HPRT and then subtracting the nucleus value and getting a fold change of gene expression compared to nucleus ( N = 3). (F) RNA scope assay showing the spatial expression of linc-ADAIN (red) and nuclei (DAPI/blue) in scramble and linc-ADAIN shRNA hTERT ASC adipocytes (scale bar, 20 μm). (G) UMAP (uniform manifold approximation and projection) projection of linc-ADAIN (linc01230) expression in single-cell RNA-seq of human subcutaneous WAT (Broad Institute ). Data are presented as the mean ± SEM.

Article Snippet: The expression plasmid was linearized and transcribed in vitro using AmpliScribe T7 Flash Biotin RNA Transcription Kit (Lucigen ASB71110) following the manufacturer’s instructions.

Techniques: RNA Sequencing, Binding Assay, Modification, Expressing, In Vitro, Cell Fractionation, Gene Expression, RNAscope, shRNA

Journal: Cell reports

Article Title: linc-ADAIN , a human adipose lincRNA, regulates adipogenesis by modulating KLF5 and IL-8 mRNA stability

doi: 10.1016/j.celrep.2024.114240

Figure Lengend Snippet: (A–F) RNA immunoprecipitation (RIP) assays measure the interaction of IL-8, KLF5 , and GAPDH with HuR (A, C, E) ( N = 3) and IGF2PBP2 (B, D, F) ( N = 3–4), upon linc-ADAIN shRNA knockdown compared to scramble shRNA in day 14 ASC hTERT adipocytes. (G–I) Scramble and linc-ADAIN shRNA-expressing ASC adipocytes were treated with actinomycin-D to halt transcription and fraction of mRNA measured at 0.5, 1, 2, and 4 h post treatment of IL-8 (normalized to GAPDH) (G), KLF5 (normalized to GAPDH) (H), and GAPDH (I) ( N = 3). * p < 0.05, ** p < 0.01 **** p < 0.0001 with respect to scramble shRNA by two-way ANOVA. Data are presented as the mean ± SEM.

Article Snippet: The expression plasmid was linearized and transcribed in vitro using AmpliScribe T7 Flash Biotin RNA Transcription Kit (Lucigen ASB71110) following the manufacturer’s instructions.

Techniques: RNA Immunoprecipitation, shRNA, Knockdown, Expressing

Journal: Cell reports

Article Title: linc-ADAIN , a human adipose lincRNA, regulates adipogenesis by modulating KLF5 and IL-8 mRNA stability

doi: 10.1016/j.celrep.2024.114240

Figure Lengend Snippet:

Article Snippet: The expression plasmid was linearized and transcribed in vitro using AmpliScribe T7 Flash Biotin RNA Transcription Kit (Lucigen ASB71110) following the manufacturer’s instructions.

Techniques: Virus, Plasmid Preparation, Recombinant, Gene Expression, shRNA, Mass Spectrometry, Negative Control, Software, Modification, Magnetic Beads, RNAscope, Multiplex Assay, Western Blot