animal-free recombinant human vitronectin  (PeproTech)

Journal: Advanced Science

Article Title: The Role of CAF‐derived Vitronectin in Promoting Colorectal Cancer Progression and Immunosuppression

doi: 10.1002/advs.202505769

Figure Lengend Snippet: Vitronectin Significantly Up‐regulated in CAFs Correlates with Poor Clinical Outcomes in CRC Patients A) Venn diagram demonstrating the intersection of upregulated differentially expressed genes (DEGs) in cancer‐associated fibroblasts (CAFs) compared to normal fibroblasts (NFs), as identified in GSE51257 , GSE92945 , and GSE231559 datasets. B) Heatmap showing the expression patterns of the overlapping genes across the three datasets. C) Violin plots illustrating the expression levels of fibroblast marker genes (ACTA2, COL1A1, COL1A2) and VTN within each cluster, based on a single‐cell RNA sequencing dataset ( GSE231559 ) comprising 6 CRC tissues and 3 adjacent normal tissues. D) tSNE plot of total fibroblasts isolated and re‐clustered from GSE231559 , color‐coded by sample origin (left) and VTN expression (right). E) Representative images of immunofluorescence costaining for VTN (red) with cell‐type markers (green) in CRC tissues from patients. Scale bars: 50 µm. F) Representative images of immunofluorescence co‐staining for VTN (red) with Vimentin, S100A4, and FAP (green) in CRC tissues from patients. Scale bars: 100 µm. G) Representative immunohistochemical staining of VTN in CRC tissue microarray. Magnification: 10X (left) and 200X (right). P: Parenchyma; S: Stroma. H) Immunohistochemical Scoring of VTN in CRC tissues and adjacent normal tissues. Student's t‐test. ***p<0.001. I) Classification of CRC and adjacent tissues into high and low VTN expression groups, followed by Chi‐Square test analysis. p<0.0001, χ2 = 26.97. J) Kaplan‐Meier survival analysis of the impact of VTN expression intensity on progression‐free survival (PFS, left) and overall survival (OS, right) of CRC patients. n = 33 patients for VTN‐low group; n = 47 patients for VTN‐high group. Log‐rank test. p<0.001. K) Higher VTN expression is associated with poorer PFS in The Cancer Genome Atlas (TCGA) CRC cohort (left), and increased distant metastasis rates in the GSE72970 CRC cohort (right). L,M) Verification of CAFs and NFs isolated from CRC tissues and paired noncancerous tissues by immunofluorescent staining (L) and western blot (M) for α‐SMA, Vimentin, FAP, and VTN, with higher expression observed in CAFs compared to NFs. Scale bar: 100 µm. N) qRT‐PCR analysis of VTN mRNA levels in 34 pairs of NFs and CAFs isolated from CRC patients. Paired two‐tailed Student's t‐test was used to calculate p value. P) ELISA detection of VTN expression in plasma samples from CRC patients (n = 36) and healthy individuals (n = 35). Student's t‐test was used to calculate p value. CAFs, cancer‐associated fibroblasts; NF, normal fibroblasts; t‐SNE, t‐distributed stochastic neighbor embedding; CRC, colorectal cancer.

Article Snippet: The impact of different cell supernatants or recombinant human vitronectin (rhVTN, MedChemExpress, USA) on oxaliplatin‐induced apoptosis in RKO and DLD1 cells was assessed using the annexin V‐FITC/propidium iodide (PI) apoptosis detection kit (Dojindo, Japan) followed by flow cytometry.

Techniques: Expressing, Marker, RNA Sequencing, Isolation, Immunofluorescence, Staining, Immunohistochemical staining, Microarray, Western Blot, Quantitative RT-PCR, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: iScience

Article Title: Enhancing human pluripotent stem cell differentiation to cardiomyocytes through cardiac progenitor reseeding and cryopreservation

doi: 10.1016/j.isci.2025.112452

Figure Lengend Snippet: EOMES + mesoderm and ISL1 +/ NKX2-5 + cardiac progenitors can be cryopreserved and differentiated on defined extracellular matrix proteins (A) Flow cytometry analysis of cTnT expression in hPSC-CMs for control (CTRL) differentiation samples compared to cryopreserved samples plated onto Fibronectin (FN), Laminin-111 (LAM1), Matrigel (MG), or Vitronectin (VTN) at either D2 or D5 from the WTC11 iPSC line. Points represent the mean values of 2–4 technical replicates for 9 independent differentiations. (B) Flow cytometry analysis of cTnT expression in hPSC-CMs for control (CTRL) differentiation samples compared to cryopreserved samples plated onto Fibronectin (FN), Laminin-111 (LAM1), Matrigel (MG), or Vitronectin (VTN) at either D3 or D6 from GiWi cell lines. Points represent the mean values of 3–4 technical replicates for 3 independent differentiations (2 IMR90-4, 1 19-9-11). (C and E) Contractile parameters (beats per minute – C and contraction duration – E in CMs differentiated from the WTC11 iPSC line quantified using MUSCLEMOTION. Points represent the mean values of 1–4 technical replicates for 8 independent differentiations, which are represented by unique symbols. (D and F) Contractile parameters (beats per minute – D and contraction duration – F in CMs differentiated from GiWi cell lines quantified using MUSCLEMOTION. Points represent the mean values of 1–4 technical replicates for 3 independent differentiations, which are represented by unique symbols (2 IMR90-4, 1 19-9-11). All p -values from two-factor linear mixed-effects model with repeated measures testing for a main column effect with a Dunnett’s post-hoc test. ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, ∗∗∗∗ for p < 0.0001, and ns for p > 0.05. All data are represented as mean ± SD.

Article Snippet: Animal-Free Recombinant Human Vitronectin , PeproTech , Cat#AF14009.

Techniques: Flow Cytometry, Expressing, Control

Journal: iScience

Article Title: Enhancing human pluripotent stem cell differentiation to cardiomyocytes through cardiac progenitor reseeding and cryopreservation

doi: 10.1016/j.isci.2025.112452

Figure Lengend Snippet: EOMES + mesoderm and ISL1 +/ NKX2-5 + cardiac progenitors can be cryopreserved and differentiated on defined extracellular matrix proteins (A) Flow cytometry analysis of cTnT expression in hPSC-CMs for control (CTRL) differentiation samples compared to cryopreserved samples plated onto Fibronectin (FN), Laminin-111 (LAM1), Matrigel (MG), or Vitronectin (VTN) at either D2 or D5 from the WTC11 iPSC line. Points represent the mean values of 2–4 technical replicates for 9 independent differentiations. (B) Flow cytometry analysis of cTnT expression in hPSC-CMs for control (CTRL) differentiation samples compared to cryopreserved samples plated onto Fibronectin (FN), Laminin-111 (LAM1), Matrigel (MG), or Vitronectin (VTN) at either D3 or D6 from GiWi cell lines. Points represent the mean values of 3–4 technical replicates for 3 independent differentiations (2 IMR90-4, 1 19-9-11). (C and E) Contractile parameters (beats per minute – C and contraction duration – E in CMs differentiated from the WTC11 iPSC line quantified using MUSCLEMOTION. Points represent the mean values of 1–4 technical replicates for 8 independent differentiations, which are represented by unique symbols. (D and F) Contractile parameters (beats per minute – D and contraction duration – F in CMs differentiated from GiWi cell lines quantified using MUSCLEMOTION. Points represent the mean values of 1–4 technical replicates for 3 independent differentiations, which are represented by unique symbols (2 IMR90-4, 1 19-9-11). All p -values from two-factor linear mixed-effects model with repeated measures testing for a main column effect with a Dunnett’s post-hoc test. ∗ for p < 0.05, ∗∗ for p < 0.01, ∗∗∗ for p < 0.001, ∗∗∗∗ for p < 0.0001, and ns for p > 0.05. All data are represented as mean ± SD.

Article Snippet: Animal-Free Recombinant Human Vitronectin (PeproTech - AF14009) was resuspended in DPBS (ThermoFisher - 14190144) and plates were coated at 3.8 μg/cm 2 .

Techniques: Flow Cytometry, Expressing, Control