Journal: bioRxiv

Article Title: Patient-Derived Circulating Monocytes Promote Calcific Aortic Valve Disease Progression

doi: 10.64898/2026.04.30.721898

Figure Lengend Snippet: PBMCs were stained for an 11-color panel across 55 samples (NCAVD, n = 20; CAVD, n = 15; volunteers, n = 20). Each panel included lineage markers CD20, CD3, CD4, CD8, and CD25 to exclude B and T lymphocytes, and monocyte markers CD14 and CD16 to identify monocyte subsets. Additional markers CD86, CD163, and CD206 were used to characterize monocyte polarization toward unpolarized, pro-inflammatory, or immunomodulatory macrophage phenotypes, respectively. The gating strategy for identifying these populations is shown for each group.

Article Snippet: CD14 + monocyte RNA samples extracted from volunteers (Vol, n=5), NCAVD (n=5), and CAVD (n=5) patients, with at least 400 ng of total RNA, with a concentration >20 ng/ml and a RIN ratio >8, were sent to Novogene Company Ltd (Cambridge, UK) for library preparation, mRNA sequencing, and bioinformatic analysis.

Techniques: Staining

Journal: bioRxiv

Article Title: Patient-Derived Circulating Monocytes Promote Calcific Aortic Valve Disease Progression

doi: 10.64898/2026.04.30.721898

Figure Lengend Snippet: IL4 and CCL3 mRNA expression were quantified by RT-qPCR (A) and normalized to the housekeeping gene B2M . Sample sizes were as follows: for IL4 , n = 8 per group; for CCL3 , n = 8 in the NCAVD and healthy volunteers (Vol) groups and n = 6 for CAVD group. (B) IL-4 and CCL3 CD14 + monocytes secretion were quantified by multiplex immunoassays. Sample sizes were as follows: for IL-4, n = 6 per group; for CCL3, n = 6 in the healthy volunteers (Vol) group and n = 5 for NCAVD and CAVD groups. Asterisks (*) indicate statistically significant differences compared to the healthy volunteer condition (blue) as determined by One-Way ANOVA followed by Sidak’s post-hoc test. Outliers were removed. p-values are indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: CD14 + monocyte RNA samples extracted from volunteers (Vol, n=5), NCAVD (n=5), and CAVD (n=5) patients, with at least 400 ng of total RNA, with a concentration >20 ng/ml and a RIN ratio >8, were sent to Novogene Company Ltd (Cambridge, UK) for library preparation, mRNA sequencing, and bioinformatic analysis.

Techniques: Expressing, Quantitative RT-PCR, Multiplex Assay

Journal: bioRxiv

Article Title: Patient-Derived Circulating Monocytes Promote Calcific Aortic Valve Disease Progression

doi: 10.64898/2026.04.30.721898

Figure Lengend Snippet: The secretome of CAVD CD14 + monocytes is depleted in cytokines involved in T lymphocyte recruitment (CXCL9, CCL21) and Th2 response promotion (CCL17, CCL22). Data are normalized to the housekeeping gene B2M. Asterisks (*) indicate statistically significant differences compared to healthy volunteer samples (blue), determined by one-way ANOVA followed by Sidak’s post-hoc test. Outliers were removed. Statistical significance is denoted as *p < 0.05 and ***p < 0.001.

Article Snippet: CD14 + monocyte RNA samples extracted from volunteers (Vol, n=5), NCAVD (n=5), and CAVD (n=5) patients, with at least 400 ng of total RNA, with a concentration >20 ng/ml and a RIN ratio >8, were sent to Novogene Company Ltd (Cambridge, UK) for library preparation, mRNA sequencing, and bioinformatic analysis.

Techniques: