Journal: The Journal of Clinical Investigation

Article Title: Tet2-driven clonal hematopoiesis drives aortic aneurysm via macrophage-to-osteoclast–like differentiation

doi: 10.1172/JCI198708

Figure Lengend Snippet: ( A ) Schematic of flow cytometric analysis and quantification of immune cells in abdominal aortic tissue from the AAA model after 1 week of AngII infusion. ( B ) Representative flow cytometry plots of digested abdominal aortic tissue 1 week after AngII infusion ( n = 5 mice per group). ( C ) Quantification of immune cell populations in digested abdominal aortic tissue (absolute number per 100 mg). Neut, neutrophils; Ly6C hi Mono, Ly6C hi monocytes; Mac, macrophages; CCR2 + Mac, CCR2 + macrophages. Saline, n = 6; AngII, n = 5; B cells and T cells, n = 5 per group. * P < 0.05 and ** P < 0.01, by Mann-Whitney U test. ( D ) Representative images of CD68, α–smooth muscle actin (α-SMA), CD31/platelet endothelial cell adhesion molecule 1 (PECAM-1), and DAPI immunofluorescence staining of abdominal aortic tissue from aortas of control and 20% Tet2 -KO BM recipient mice after 1 week of AngII infusion. Images are representative of 8 mice per genotype. Scale bars: 100 μm. ( E ) Schematic of RNA-seq analysis of sorted macrophages from abdominal aortas of 100% Tet2 -KO BM recipient mice after 1 week of AngII infusion. ( F and G ) Upregulated and downregulated genes are presented as a heatmap ( F ) and volcano plot ( G ). ( H ) Gene Ontology enrichment analysis of upregulated genes in Tet2 -deficient macrophages. The table lists the predominant genes ranked by q value among significantly enriched pathways. ( I and J ) In vitro differentiation of BM-derived macrophages. ( I and J ) Representative images and quantification of the TRAP + cell-to-macrophage ratio after RANKL stimulation from 8 independent biological replicates per genotype. *** P < 0.001, by Mann-Whitney U test. ( K ) qRT-PCR analysis of BM-derived macrophages 6 hours after stimulation with 10 ng/mL LPS ( n = 6 independent biological replicates per genotype). **** P < 0.0001, by Mann-Whitney U test.

Article Snippet: Lentiviral titers were determined using a Lenti-X quantitative reverse transcription PCR (qRT-PCR) Titration Kit (catalog 631235, Clontech).

Techniques: Flow Cytometry, Saline, MANN-WHITNEY, Immunofluorescence, Staining, Control, RNA Sequencing, In Vitro, Derivative Assay, Quantitative RT-PCR

Journal: Renal Failure

Article Title: Inhibition of caspase-1 by ginsenoside Rg1 ameliorates d -gal-induced renal aging and injury through suppression of oxidative stress and inflammation

doi: 10.1080/0886022X.2025.2504634

Figure Lengend Snippet: Changes of caspase-1 signal pathway in kidney tissue after Rg1 intervention. (A) qRT-PCR analysis of caspase-1 transcription in kidney in each group. (B) qRT-PCR analysis of IL-1 transcription in kidney in each group. (C) qRT-PCR analysis of IL-18 transcription in kidney in each group. (D) Western blots of caspase-1 of kidney tissue. (E) Relative protein expression of caspase-1 in kidney. (F) The level of interleukin-1 (IL-1, pg) in kidney measured by ELISA kit. (G) The level of interleukin-18 (IL-18, ng) in kidney measured by ELISA kit. The data are expressed as the means ± SEM, n = 3. * p < 0.05, ** p < 0.01, ** p < 0.001.

Article Snippet: PrimeScript ™ RT reagent kit (RR037A) and TB Green ® Premix Ex Taq ™ II (RR820A) used for quantitative reverse transcription PCR (qRT-PCR) experiments were purchased from TAKARA (Kusatsu, Shiga, Japan).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Direct regulation of Per2 by Roraa: insights into circadian and metabolic interplay in zebrafish

doi: 10.1007/s00018-025-05696-8

Figure Lengend Snippet: Circadian expression of the roraa gene in zebrafish is regulated by the circadian clock. A Under LD (light/dark) conditions, qRT‒PCR was used to measure the mRNA expression levels of the roraa gene in wild-type (WT) zebrafish. JTK cycle analysis revealed that roraa expression exhibited significant circadian oscillation (p < 0.05). B In situ hybridization was performed to detect roraa gene expression under LD conditions, revealing that roraa expression during the day was significantly greater than that at night, demonstrating circadian rhythmic differences. C Statistical results from in situ hybridization under LD conditions revealed that roraa expression was significantly higher during the day than at night. D Potential cis-regulatory elements on the roraa promoter fragment were predicted via the JASPAR database; two D-box elements, two E-box elements, and one RORE element, which may play key regulatory roles in the circadian expression of roraa , were identified. E Dual-luciferase reporter assays revealed that the heterodimers formed by the Bmal1b and Clocka proteins, as well as the Tefa, Roraa, Rorb, and Rorc proteins, significantly activated the expression of the roraa gene. F Diagram showing the deletion of E-box elements in the roraa -luc plasmid. G The results from dual-luciferase reporter assays demonstrated that the activation of the roraa gene by Bmal1b and Clocka was significantly reduced when E-box1 and E-box2 elements were deleted from the roraa -luc plasmid, indicating the important role of E-box elements in the regulation of roraa expression. All the data are presented as the means ± standard errors of the means (SEMs) (n = 3); ** indicates p < 0.01, **** indicates p < 0.0001 (Student's t test)

Article Snippet: The extracted RNA was reverse transcribed into cDNA via a qRT‒PCR reverse transcription kit (Vazyme, Code No:R423-01).

Techniques: Expressing, In Situ Hybridization, Gene Expression, Luciferase, Plasmid Preparation, Activation Assay