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: Oncogene

Article Title: Wnt target IQGAP3 promotes Wnt signaling via disrupting Axin1-CK1α interaction

doi: 10.1038/s41388-025-03512-y

Figure Lengend Snippet: A Immunoblot of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. Samples were subjected to SDS-PAGE (7.5%) followed by immunoblotting using the indicated antibodies. The relative immunoblot bands of β-catenin and β-actin (βcat/βact) were quantified by densitometry. B NUGC3 cells were lysed, and RNA collected were converted to cDNA and subjected to RT-PCR to quantify the amount of Wnt target genes CCND1 and MYC . Representative data were collected and are expressed as the mean ± SD from three independent experiments. Student’s t test was performed, with ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001. C XTT Cell Proliferation Assay of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. Representative data were collected and are expressed as the mean % growth ±SD from three independent experiments. Two-way ANOVA was performed, with ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001. D Clonogenic Assay of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. E Colony-Forming Efficiency (%) of NUGC3 wild-type and IQGAP3 CRISPR knock-out clones. F Immunoblot of AGS wild-type and IQGAP3 CRISPR knock-out clones. Image is best representative of three independent experiments. Samples were subjected to SDS-PAGE (7.5%) followed by immunoblotting using the indicated antibodies. The relative immunoblot bands of β-catenin and α-Tubulin (βcat/αTub) were quantified by densitometry. G AGS cells were lysed, and RNA collected were converted to cDNA and subjected to RT-PCR to quantify the amount of Wnt target genes CCND1 and MYC . Representative data were collected and are expressed as the mean ± SD from three independent experiments. Student’s t test was performed, with ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001. H XTT Cell Proliferation Assay of AGS wild-type and IQGAP3 CRISPR knock-out clones. Representative data were collected and are expressed as the mean % growth ±SD from three independent experiments. Two-way ANOVA was performed, with ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001. I Clonogenic Assay of AGS wild-type and IQGAP3 CRISPR knock-out clones. J Colony-Forming Efficiency (%) of AGS wild-type and IQGAP3 CRISPR knock-out clones.

Article Snippet: Cell proliferation was determined using the TACS® XTT Cell Proliferation Assay (R&D Systems, Inc.), following the manufacturer’s instructions.

Techniques: Western Blot, CRISPR, Knock-Out, Clone Assay, SDS Page, Reverse Transcription Polymerase Chain Reaction, Proliferation Assay, Clonogenic Assay