lna-modified oligonucleotide probe  (Enzo Biochem)

Journal: Theranostics

Article Title: Hyperglycemia facilitates EV71 replication: Insights into miR-206-mediated regulation of G3BP2 promoting EV71 IRES activity

doi: 10.7150/thno.93883

Figure Lengend Snippet: miR-206 exhibits significant impacts on EV71 neurovirulence and insulin secretion in hSCARB2-Tg mice. (A) qRT-PCR analysis of viral load in the brainstem of EV71-infected hSCARB2-Tg mice, with or without antagomiR-206 (AM206) treatment (n = 5). Statistical significance was determined using a two-sided T-test, with a significance threshold set at p < 0.05. (B) Survival assessment of EV71-infected hSCARB2-Tg mice, with or without AM206 treatment. Seven-day-old hSCARB2-Tg mice infected with EV71 were administered LNA miR-206 antagomir (AM206, 1.2 mg/kg, n = 15) or (Ctrl, n = 14). The log-rank test was utilized to determine the statistical significance in survival rates between the treatment groups. (C) Representative micrographs depicting brainstem gliosis in EV71-infected hSCARB2-Tg mice, with or without AM206 treatment. Scale bar represents 20 μm. (D) Brainstem expression of proinflammatory cytokines in EV71-infected hSCARB2-Tg mice, with or without AM206 treatment. RNA was extracted from the brainstem tissue of mice at 5 dpi and subjected to qRT-PCR analysis using TNFα-specific, and IL-6-specific primers, normalized to GAPDH (n = 5). (E) Proposed role of miR-206 in targeting GCK-mediated insulin-glucose homeostasis. (F) Representative immunohistochemistry (IF) images displaying GCK levels in the pancreas of EV71-infected hSCARB2-Tg mice, with or without AM206 treatment. Paraffin-embedded sections were stained using the VP1/2 antibody to detect EV71, while islet staining was performed using an insulin antibody. Scale bar denotes 20 μm. (G) qRT-PCR quantification of GCK levels in the pancreas of EV71-infected hSCARB2-Tg mice, with or without AM206 treatment (n = 5). (H) Plasma insulin levels in EV71-infected hSCARB2-Tg mice, with or without AM206 treatment. Insulin levels were measured using the mouse Insulin ELISA assay (n = 5). (I) Plasma glucose levels in EV71-infected mice, with or without AM206 treatment. Glucose levels were measured using a blood glucose assay (n = 5).

Article Snippet: LNA-modified microRNA miR-206 antisense oligonucleotide probes were labeled with Biotin (MDBio Inc.) and the signal was amplified using Tyramide amplification solution (Thermo Fisher Scientific).

Techniques: Quantitative RT-PCR, Infection, Expressing, Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay, Glucose Assay

Journal: Genome Biology

Article Title: Altered retinal microRNA expression profile in a mouse model of retinitis pigmentosa

doi: 10.1186/gb-2007-8-11-r248

Figure Lengend Snippet: miR ISH analysis in the mouse retina. Eyes from 1-month-old c57 animals were fixed in 4% paraformaldehyde, and 12 μm cryosections were in situ hybridized with 5'-digoxigenin labeled locked nucleic acid (LNA) microRNA (miR) probes for (a) let-7, (b) miR-181a, and (c,d) miR-182. A false-colored (magenta) 4',6-diamidine-2-phenylindole-dihydrochloride (DAPI) nuclear staining is overlaid on the miR-182 in situ hybridization (ISH) label (panel d) to indicate the position of the nuclear layers. Scale bar: 25 μm. GCL, ganglion cell layer; INL, inner nuclear layer; IS, photoreceptor inner segments; ONL, outer nuclear layer; OS, photoreceptor outer segments.

Article Snippet: 5'-Digoxigenin (DIG) labeled, LNA-modified oligonucleotide ISH probes were purchased from Exiqon for the following mouse miRs: 1, 9*, 26b, 96, 129-3p, 133, 138, 181a, 182 and 335, and let-7d (including sense-159) as background control.

Techniques: In Situ, Labeling, Staining, In Situ Hybridization