Journal: Cell reports

Article Title: An enhancer RNA recruits KMT2A to regulate transcription of Myb

doi: 10.1016/j.celrep.2024.114378

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RNA pulldown was performed using the Pierce Magnetic RNA-Protein Pull-Down Kit (ThermoFisher Scientific, 20164) according to the manufacturer’s protocol.

Techniques: Recombinant, SYBR Green Assay, Electroporation, Transfection, Protease Inhibitor, Magnetic Beads, Gel Extraction, Cell Isolation, Plasmid Preparation, Software

Journal: Epigenetics

Article Title: Circ-0006332 stimulates cardiomyocyte pyroptosis via the miR-143/TLR2 axis to promote doxorubicin-induced cardiac damage

doi: 10.1080/15592294.2024.2380145

Figure Lengend Snippet: Circ -0,006,332 overexpression accelerates DOX-induced cardiomyocyte pyroptosis through miR-143 downregulation. a: RT-qPCR to detect miR-143 and circ -0,006,332 expression in rat heart tissues; b: the FISH assay to detect intracellular co-localization of circ -0,006,332 and miR-143 in cardiomyocytes; c: dual-luciferase assay to detect the binding relationship between miR-143 and circ -0,006,332 in H9C2 cells; d: circ 0006332 and miR-143 expression measured by RT-qPCR. e: RNA pull-down assay to detect the binding of miR-143 to circ -0,006,332; f: CCK-8 assay to detect cardiomyocyte viability; g: LDH release in cardiomyocyte supernatants; h: intracellular ATP contents; i-k: ELISA to detect IL-1β, IL-18, and TNF-α levels in cell supernatants; l-m: protein levels of TLR2 and pyroptosis-related molecules (ASC, cleave-caspase-1, GSDMD-N, NLRP3, and HMGB1) in cardiomyocytes detected by western blotting. In panel A, * p < 0.05 compared with the vehicle group; in panel C, * p < 0.05 compared with the WT + mimic NC group; in panel D, * p < 0.05 compared with the IgG group; in panels F-M, * p < 0.05 compared with the vehicle + mimic NC + oe-NC group, # P < 0.05 compared with the DOX + mimic NC + oe-NC group, and $ p < 0.05 compared with the DOX+ miR-143 mimic + oe-NC group. Values in the figures were measurement data and expressed as mean ± standard deviation. Two-group comparisons were analysed with the independent sample t-test (A, D, E), and one-way ANOVA with Tukey’s post hoc test (C-F-K, M) was used for comparisons among multiple groups. The cell experiments were repeated three times.

Article Snippet: Based on the manuals of the Pierce TM Magnetic RNA-Protein Pull-Down kit (Millipore), the biotinylated miR-143 probe or control probe (Geneseed, Guangzhou, China) was incubated with cell lysate at 25°C for 2 h. The complexes were captured with streptavidin-labelled immunomagnetic beads and incubated with proteinase K-contained buffer for 1 h. Eluted complexes were measured with RT-qPCR [ ].

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Luciferase, Binding Assay, Pull Down Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation