Journal: Journal of Hematology & Oncology

Article Title: Super-enhancer hijacking LINC01977 promotes malignancy of early-stage lung adenocarcinoma addicted to the canonical TGF-β/SMAD3 pathway

doi: 10.1186/s13045-022-01331-2

Figure Lengend Snippet: Epigenetic transcriptional activation of the SMAD3/CBP/P300 complex is LINC01977 -dependent and can potentially serve as a target for combination therapy. A RNA-binding abilities of LINC01977 and CBP/P300 were predicted by OmiXcore. B RNA pull-down and western blot assays were used to evaluate the interaction between LINC01977 and CBP/P300. C RIP and qRT-PCR assays were used to measure the enrichment of LINC01977 . D Co-immunoprecipitation (co-IP) of CBP or P300 with phosphorylated SMAD3 Ser423/425 in A549 cells transfected with pcDNA3.1 or LINC01977 followed by treatment with TGF-β (10 ng/mL). E Co-IP assays of CBP or P300 with phosphorylated SMAD3 Ser423/425 in A549 cells, which were transfected with LINC01977 followed by treatment with TGF-β (10 ng/mL), and treated with RNase inhibitor or RNase. F Represent images of H3K27ac, assayed by IF confocal microscopy in A549 cells treated with TGF-β (10 ng/mL). Actin, Phalloidin (red); nuclei, DAPI (blue); and H3K27ac (green). Scale bar: 20 μm. ( G, H ) ChIP assays suggested that LINC01977 SE ( G ) and promoter ( H ) were co-occupied by CBP, P300 and H3K27ac. I Abilities of invasion and migration were diminished by combination of LINC01977 -ASO and inhibition of CBP/P300, SGC-CBP30 (5 μM) in vitro. J Schematic representation of lung metastasis nude mouse model. K Representative images of IVIS detection and metastatic loci in the lung derived from mouse model. L Mouse overall survival after tail-vein injection. M H&E stained and phosphorylated SMAD3 Ser423/425, CBP, and ZEB1 IHC detection of serial sections from lung metastasis nude mouse model. Representative images are shown. H&E, scale bars: 500 μm; IHC, scale bars: 50 μm. Data in C, G–I are representative of three independent experiments and presented as mean ± S.D., n = 3 biologically independent samples, and the P value was determined by a two-tailed unpaired Student’s t test and one-way ANOVA, respectively. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; n.s. not significantly

Article Snippet: RIP assays were performed with the Magna Nuclear RIP (Native) Nuclear RNA-Binding Protein Immunoprecipitation Kit (Merck Millipore) according to the manufacturer’s instructions.

Techniques: Activation Assay, RNA Binding Assay, Western Blot, Quantitative RT-PCR, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Confocal Microscopy, Migration, Inhibition, In Vitro, Derivative Assay, Injection, Staining, Two Tailed Test

Journal: BioMed Research International

Article Title: LncRNA PVT1 Promotes Hypoxia-Induced Cardiomyocyte Injury by Inhibiting miR-214-3p

doi: 10.1155/2021/4604883

Figure Lengend Snippet: Relationship between PVT1 and miR-214-3p. (a) DLR confirmed that there was a binding relationship between PVT1 and miR-214-3p. (b) Enrichment ability of PVT1 and AGO2 evaluated by RIP assay. (c) Enrichment ability of PVT1 and miR-214-3p determined by RNA pull-down. ∗ indicated P < 0.05.

Article Snippet: As instructed by the guidelines, EZ-Magna RIP kit (Shanghai Advantage Biological Co., Ltd.) was utilized for RNA immunoprecipitation.

Techniques: Binding Assay