Journal: International Journal of Molecular Sciences

Article Title: Thrombospondin 1–CD47 Signalling Modulates Vascular Smooth Muscle Cell Senescence in Chronic Kidney Disease

doi: 10.3390/ijms27020755

Figure Lengend Snippet: TSP1 and indoxyl sulfate limit VSMC proliferation via CD47. hVSMC cell viability was measured by assessing reduction in tetrazolium salt sodium 3′- [1- [(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT) and measuring absorbance at 450 nm in cells after 48 h treatment with ( A ) TSP1 (0, 0.2, 2.2, 5, 10 nM) ( n = 4), ( B ) TSP1 2.2 nM ± pre-treatment with anti-CD47 antibody for 30 min ( n = 6), ( C ) IS (0, 1, 10, 100, 500 µM) ( n = 4), or ( D ) IS (100, 500 µM) ± pre-treatment with anti-CD47 antibody for 30 min ( n = 4–8). All data shown are mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 by one-way analysis of variance with Holm–Sidak post-hoc test ( A , C ) or Kruskal–Wallis test ( B , D ). Abbreviations: αCD47—anti-CD47 antibody; hVSMC—human aortic vascular smooth muscle cell; IS—indoxyl sulfate; TSP1—thrombospondin-1.

Article Snippet: Cell proliferation and senescence were measured using XTT Cell Viability Kit (#9095, Cell Signalling Technology, Danvers, MA, USA) and Mammalian β-Galactosidase Assay Kit (75707, Thermo Fisher Scientific), respectively.

Techniques:

Journal: Scientific Reports

Article Title: EMID1, a multifunctional molecule identified in a murine model for the invasion independent metastasis pathway

doi: 10.1038/s41598-021-96006-2

Figure Lengend Snippet: Effect of EMID1 expression on cell properties in vitro. ( A ) Cell proliferation analysis using the XTT assay. 66–4 cells stably overexpressing EMID1 and 66HM cells transfected with siRNA at 48 h were measured. Error bars, ± SD (n = 3). ** P < 0.01. ( B ) Cell cycle analysis using flow cytometry. 66HM cells transfected with siE2 at 24 h were analyzed. Error bars, ± SEM (n = 3). * P < 0.05. ( C ) Cell migration analysis using the wound healing assay. The speed of wound closure for 24 h of both overexpressing and knockdown cells was evaluated (upper). Error bars, ± SD (n = 3). ** P < 0.01. Phase contrast images of migrating cells (bottom). Scale bar, 500 μm. ( D ) Cell invasion analysis using the biocoat matrigel chamber. After incubation of 66–4 and 66HM transfectants for 8 and 12 h, respectively, migrating cells were counted (upper). Error bars, ± SD (n = 3). * P < 0.05. Representative image of invading cells stained with hematoxylin (bottom). Scale bar, 100 μm.

Article Snippet: The effect of EMID1 overexpression or knockdown on in vitro cell growth was examined using an XTT assay kit (Merck).

Techniques: Expressing, In Vitro, XTT Assay, Stable Transfection, Transfection, Cell Cycle Assay, Flow Cytometry, Migration, Wound Healing Assay, Incubation, Staining