Journal: The FASEB Journal

Article Title: Loss of MFAP5 and Its Effect on Skin Homeostasis and Wound Healing

doi: 10.1096/fj.202501770R

Figure Lengend Snippet: Mfap5 −/− fibroblasts retain their inhibited migratory and proliferative capacity across different ECM substrates. In vitro vertical wounds after one‐by‐one cross‐scratching for Mfap5 +/+ and Mfap5 −/− fibroblast migration assay on fibronectin (A) coated plates expressed as a percent of the wound closed (B) at each time point. Mfap5 +/+ and Mfap5 −/− fibroblast migration assay on collagen I (C) coated plates expressed as a percent of the wound closed (D). N = 12 for B and D. Areas not covered by cells are outlined by a black line. MTS proliferation assay was performed on Mfap5 +/+ and Mfap5 −/− fibroblasts at 24‐, 48‐, and 72‐h post‐seeding onto a fibronectin (E) or collagen I (F) coated plate. N = 8 for E and F, with each dot representing a biological replicate that consists of 4–6 technical replicates. (G) Heatmap of differentially expressed matrix metalloproteinases ( Mmp ) and integrins ( Itg ) between Mfap5 +/+ (WT) and Mfap5 −/− (KO) fibroblasts. Bars on all graphs indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. Two‐way ANOVA with two‐stage linear step‐up procedure of Benjamini, Krieger, and Yekutieli post hoc testing (vs Mfap5 +/+ ).

Article Snippet: Cell migration and proliferation across different substrates were assessed by seeding Mfap5 +/+ or Mfap5 −/− fibroblasts into 12‐ and 96‐well plates coated with rat tail collagen 1 (BD Biosciences) or mouse fibronectin (Innovative Research Inc., Novi, MI), at 5 μg/cm 2 .

Techniques: In Vitro, Migration, Proliferation Assay

Journal: Aging Cell

Article Title: A new gene signature for endothelial senescence identifies self‐ RNA sensing by retinoic acid‐inducible gene I as a molecular facilitator of vascular aging

doi: 10.1111/acel.14240

Figure Lengend Snippet: Establishing and benchmarking a senescence gene signature for endothelial cells. (a) Images of early passage (EP) and senescent (Sen) ECFCs stained for senescence‐ associated β‐Galactosidase activity (SA‐β‐Gal). Scale bar: 50 μm. Representative cell culture wells stained with crystal violet to assess for clonogenic capacity. Scale bar: 5 mm. 3D Matrigel angiogenesis assay. ECFCs were stained with calcein for fluorescent microscope imaging at Day 2. Scale bar: 100 μm. (b) Genome‐wide transcriptomics differential gene expression analysis shown in volcano plots. Significantly upregulated transcripts highlighted in purple, orange, and green. (c) Euler diagram for upregulated genes from the three cellular senescence models to identify a common senescence signature in endothelial cells. (d) Heatmap and unbiased clustering analysis for the top 20 upregulated genes based on log 2‐fold change. IFN‐related transcripts are highlighted in red. (e) The images of EP, replicative (Rep) Sen, and etoposide (Eto)‐induced Sen; human retinal microvascular endothelial cells (HRMECs) stained for SA‐β‐Gal. Scale bar: 100 μm. Heatmap and unbiased clustering analysis, using the EndoSEN_up gene signature, showing the top 10 upregulated genes in Sen HRMECs compared to EP HRMECs. (f) Normalized enrichment score (NES) for GSEA benchmarking our EndoSEN signatures to previously established senescence gene signatures. Signatures were assessed across datasets for endothelial cells (yellow background) and fibroblasts (pink background). ECFCs, endothelial colony forming cells; HAECs, human aortic endothelial cells; HRMECs, human retinal microvascular endothelial cells; HUVECs, human umbilical vein endothelial cells; WI‐38 and IMR‐90, human lung fibroblasts.

Article Snippet: For interferon treatment, cells were treated with 100 IU/mL Interferon‐β (R&D Systems) for 24 h. HRMECs were purchased from Innoprot and cultured on fibronectin‐coated flasks in ECM medium (Innoprot) supplemented with 5% FBS, Endothelial Cell Growth Supplements (ECGS) and penicillin/streptomycin.

Techniques: Staining, Activity Assay, Cell Culture, Angiogenesis Assay, Microscopy, Imaging, Genome Wide, Expressing