Journal: Kidney360

Article Title: The Positive Feedback Loop of Hypoxia-Inducible Factor-1 α /miR-295/Factor Inhibiting Hypoxia-Inducible Factor-1 in Hyperuricemic Nephropathy

doi: 10.34067/KID.0000001069

Figure Lengend Snippet: miR-295 is induced in renal tubules in HN mice. HN was induced by PO intraperitoneally and Ad orally administered for 2 weeks. Except for the control group, each group was intraperitoneally administered PO (350 mg/kg per day) and orally administered with Ad (70 mg/kg per day) to induce HN at 8:30 am for 14 consecutive days. Control mice were treated with normal saline. At the end of 21st day, the animals were euthanized. (A) Body weight changes in control and HN mice; (B) serum UA concentrations in each group; (C) serum CRE levels; (D) BUN levels; (E) representative images of HE staining, scale bar: 50 μ m; (F) representative images of Masson staining, scale bar: 50 μ m; (G) quantitative analysis of tubular injury score in different experimental groups; (H) quantification of Masson's trichrome–positive fibrotic area in kidney sections; (I) volcano plot of microRNA expression profiled by microarray in HN. Differentially expressed miRNAs were identified using data obtained from three biological replicates per experimental group ( n =3 mice per group, total six samples). Statistical significance was determined using the criteria of |log 2 fold change| >1.5 and P < 0.05. These thresholds were applied to ensure robust identification of meaningful expression differences. (J) qPCR analysis of miR-295 in mouse kidneys. The level of miR-295 was normalized to the level of U6 (internal loading control) of the same samples to determine the ratio with the ratio of control mice arbitrarily set as 1. (K) In situ hybridization showing miR-295 increase in kidney tissues. Representative images of double immunostaining with miR-295 and proximal tubule marker, LTL, scale bar: 50 μ m. (L) Quantitative analysis of fluorescence intensity. All the values are expressed as mean±SD ( n =6), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Ad, adenine; CRE, creatinine; HE, hematoxylin–eosin; HN, hyperuricemic nephropathy; LTL, Lotus tetragonolobus lectin; PO, potassium oxonate; UA, uric acid

Article Snippet: The Digoxigenin-labeled mmu-miR-295 locked nucleic acid (LNA) probe and Fluorescence in situ hybridization (FISH) Kit were from Servicebio (Wuhan, China). miR-295 mimic, anti-miR-295 LNA, and FIH-1 siRNA were from Ruibo (Guangzhou, China).

Techniques: Control, Saline, Staining, Expressing, Microarray, In Situ Hybridization, Double Immunostaining, Marker, Fluorescence

Journal: Molecular Biomedicine

Article Title: YTH N6-methyladenosine RNA binding protein 2 mediated m 6 A modification of circHIPK2 promotes cellular senescence and osteoarthritis progression by inhibiting autophagy

doi: 10.1186/s43556-026-00441-4

Figure Lengend Snippet: Expression pattern of circHIPK2 and its characteristics in human chondrocytes. a , b Histological analysis, preoperative Kellgren-Lawrence and OARSI grading of sequencing cartilage sample (n = 3 per group). c Heatmap of circRNA expression in OA and NA chondrocytes. d RT-qPCR analysis of circHIPK2, circGOSR2, circREPS1, circMTUS1, circNUP54, circSLC8A1, circADAMTS6, circFN1, circTNFRSF21 and circAPBB2 expression in NA and OA chondrocytes ( n = 6 per group). e Representative images of circHIPK2 expression in NA and OA chondrocytes via FISH staining, γH2AX staining, SA-β gal staining. f CircHIPK2 expression pattern in chondrocytes treated with IL-1β, Etoposide and TNF-α, assessed via RT-qPCR ( n = 6 per group). g RNA fluorescence in situ hybridization (FISH) staining of circHIPK2 in aging mice and DMM mice. h Schematic illustration depicting HIPK2 exons 2 circularization to form circHIPK2. The presence of circHIPK2 was validated by RT-qPCR, followed by Sanger sequencing, red box represents head-to-tail circHIPK2 splicing sites. i RT-qPCR analysis of circHIPK2 and HIPK2 expression in chondrocytes treated with or without RNase R ( n = 6 per group). j RT-qPCR analysis of circHIPK2 and HIPK2 expression in chondrocytes treated with actinomycin D for different durations ( n = 3 per group). k RT-qPCR analysis of circHIPK2 and HIPK2 expression in NA and OA chondrocytes ( n = 10 per group). l Representative images of FISH staining for circHIPK2 localization. m RT-qPCR analysis of circHIPK2 expression in the nuclear and cytoplasmic fractions. GAPDH served as control. ** P < 0.01, *** P < 0.001

Article Snippet: A RNA fluorescent in situ hybridization kit (cat. no. GF-003, Servicebio, China) was used to detect circHIPK2 expression in chondrocytes and knee joint sections.

Techniques: Expressing, Sequencing, Quantitative RT-PCR, Staining, Fluorescence, In Situ Hybridization, Control