Journal: bioRxiv

Article Title: Aurora A kinase activation contributes to the fibrotic phenotype in Systemic Sclerosis through primary cilia shortening

doi: 10.64898/2026.03.13.711548

Figure Lengend Snippet: (A) Representative immunofluorescence images of primary cilia in healthy control (HC), systemic sclerosis (SSc), and VEDOSS (very early SSc) dermal fibroblasts under basal conditions (CTR) or following 24 h TGFβ stimulation. Acetylated α-tubulin marks the axoneme (red) and nuclei are stained with DAPI (blue). (B) Quantification of primary cilium length in HC, SSc, and VEDOSS fibroblasts under basal conditions. n=3 donors per group, a minimum of 100 cilia were quantified per sample. (C) Time course of TGFβ-induced cilia shortening in HC and SSc fibroblasts. n=3 donors per group, a minimum of 100 cilia were quantified per sample. (D) Quantification of cilium length in HC and SSc fibroblasts subjected to a 24 h TGFβ pulse followed by washout and recovery in 0.5% serum for 24 and 48 h. n=3 experimental repeats. (E) Quantification of cilium length in HC and SSc fibroblasts after sequential serum starvation, TGFβ exposure, and two passages in 10% serum. n=3 experimental repeats. (F) Cilium length in HC and SSc fibroblasts treated with the TGFβ receptor inhibitor SD208 in the presence or absence of exogenous TGFβ. n=3 experimental repeats. (G) Representative western blot of pSMAD3, total SMAD3, and βactin in HC and SSc fibroblasts treated with TGFβ and/or SD208. n=3 donors per group. (H) Quantification of cilium length in HC and SSc fibroblasts cultured in full growth medium in the presence of DMSO (CTR) or SD208 for 2 passages. n=3 experimental repeats. All data panels were analysed by ANOVA (ns, non-significant; ** P<0.01; *** P<0.001; ****P<0.0001).

Article Snippet: Fibroblasts were treated with recombinant TGFβ1 ligand (10 ng/mL, Merck), TGFβRI inhibitor SD208 (1 μM, Selleckchem), ROCK2 inhibitor KD025 (5 μM, Cayman Chemical), HDAC6 inhibitor Tubacin (10 μM, Sigma), and AURKA inhibitor MLN8054 (10 μM, APExBIO).

Techniques: Immunofluorescence, Control, Staining, Western Blot, Cell Culture

Journal: Scientific Reports

Article Title: Biomimetic post-capillary venule expansions for leukocyte adhesion studies

doi: 10.1038/s41598-018-27566-z

Figure Lengend Snippet: Spatial statistics of leukocyte adhesion as a function of coatings. ( a ) Leukocyte adhesion distribution heatmap for all ICAM-1 containing conditions in the straight channel device (Supplemental Fig. ). Flow direction is left to right. These consist of a length 4 expansion and an extended straight section to capture relaxation of the expansion effect. Wall associated adhesion peaks immediately after the initial expansion, and again at the collision-inducing point where the expansions end. Channel center adhesion subtly increases over the length of the channel. ( b ) Mean number of adherent cells per ROI in response to factorial combinations of CXCL12, SDC4, and ICAM1 in the coating, demonstrating additive effects. Each point represents mean numbers for a single participant, N = 6 donors. Boxes indicate mean and standard error. ( c ) Sub-ROI quantification using a mask for wall-associated vs. center of channel leukocytes as a function of flow distance along the ROI, demonstrating relaxation of the expansion effect for wall-associated but not channel center leukocytes. Wall associated adhesion probability peaks immediately after the expansion, then declines across the ROI until the junction with the constriction induces further collisions between erythrocytes and leukocytes, generating a second peak of adhesion. In contrast, channel center adhesion gradually increases across the length of the channel. ( d ) Validation of the sub-ROI quantification method by quantifying across the cross section of the ROI: Wall associated adhesion is restricted to the sidewalls, whereas channel center adhesion peaks in the channel center. ( e ) Linear model fits across the ROI cross section for the relative contributions of CXCL12, SDC4, and ICAM1 to adhesion. As expected, the contribution of adhesion molecule ICAM1 is most significant in the channel center, where shear stress is highest. ( f ) Linear model fits along the ROI for coating contributions to leukocyte adhesion on the sidewall. The initial spike of adhesion at the expansion is strongly ICAM-1 dependent. ( g ) Linear model fits along the ROI for leukocyte adhesion in the center of the channel. Adhesion immediately after the expansion is dependent on all three coatings, whereas ICAM-1 becomes more dominant as the expansion effect relaxes.

Article Snippet: Recombinant human CXCL12 and recombinant human Fc-ICAM1 chimeric fusion protein were obtained from R&D Systems (Minneapolis, MN).

Techniques: Biomarker Discovery, Shear

Journal: Breast Cancer Research : BCR

Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

doi: 10.1186/bcr3053

Figure Lengend Snippet: Genes differentially regulated by IRF5 in MDA-MB-231 cells.

Article Snippet: Briefly, 100 ng/ml human recombinant CXCL12/SDF-1 (R&D Systems, Minneapolis, MN, USA) was added to 600 μl of phenol red-free DMEM medium supplemented with 10% FBS in the lower chamber.

Techniques: Expressing