Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/eLife.58688

Figure Lengend Snippet: Migration of activated PBT plated onto fresh tumor slices was analyzed in EGI-1 and MMTV-PyMT tumor model, whilst resident tumor-infiltrating T lymphocytes were analyzed in mPDAC and KPC tumor model. Illustrative images of T cell migration tracks in EGI-1, MMTV-PyMT, mPDAC, and KPC tumor models. Tumor stroma (fibronectin) in red, tumor cells (EpCAM in EGI-1, MMTV-PyMT, and KPC tumor models, CD44 in mPDAC tumor models), in blue and T cells (CD8 in mPDAC and KPC, Calcein in MMTV-PyMT, and EGI-1 tumor models) in green. Tracks are color-coded to illustrate track displacement. Scale bar = 100 µm. T cell migration speed, T cell displacement, and trajectory straightness in all tumor models. ***p - value>0.001, p-value>0.05, Student’s t-test. Results are shown as mean ± SD. Figure 5—source data 1. Source data file for .

Article Snippet: Live vibratome sections were stained with BV421 anti-mouse EpCAM (G8.8 clone; BD Biosciences), PerCP-e710 anti-mouse CD8a (53–6.7 clone, eBioscience), PE anti-podoplanin (8.1.1 clone; BioLegend), and anti-human/mouse fibronectin (HFN7.1 clone; Novus Biologicals).

Techniques: Migration

Journal: eLife

Article Title: Tumor stiffening reversion through collagen crosslinking inhibition improves T cell migration and anti-PD-1 treatment

doi: 10.7554/eLife.58688

Figure Lengend Snippet: ( A ) Representative images of Control and BAPN-treated KPC tumors. Tumor cells (EpCAM) in blue, tumor stroma (fibronectin) in red, and T cells (CD8) in green. Scale bar = 100 µm. ( B ) Quantification of CD8 + T cells per mm 2 in tumor islets and stroma regions in control and BAPN (*p<0.01, Student’s t-test, n = 2–3 mice/group). Figure 5—figure supplement 1—source data 1. Source data file for .

Article Snippet: Live vibratome sections were stained with BV421 anti-mouse EpCAM (G8.8 clone; BD Biosciences), PerCP-e710 anti-mouse CD8a (53–6.7 clone, eBioscience), PE anti-podoplanin (8.1.1 clone; BioLegend), and anti-human/mouse fibronectin (HFN7.1 clone; Novus Biologicals).

Techniques: Control

Journal: The American journal of pathology

Article Title: Overexpression of Corin Ameliorates Kidney Fibrosis through Inhibition of Wnt/β-Catenin Signaling in Mice.

doi: 10.1016/j.ajpath.2023.09.008

Figure Lengend Snippet: Figure 1 Wnt/b-catenin signaling is activated and corin expression is decreased along with the increased expression of b-catenin in mouse kidney after unilateral ureteral obstruction (UUO). A and B: Western blot analyses of kidney expression of several proteins, including Wnt1, a-smooth muscle actin (a- SMA), type I collagen, fibronectin, Snail, and plasminogen activator inhibitor-1 (PAI-1), in sham and obstructed kidneys at day 7 after UUO. Representative Western blot analysis (A) and quantitative data relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH; B) are presented. C: Representative mi- crographs using immunohistochemical staining for corin and active b-catenin in kidney tissues at days 4 and 7 after UUO. Boxed areas show magnification of images. Arrows indicate corin- or active b-cateninepositive tubular epithelial cells. Arrowheads in the enlarged boxed areas indicate positive staining. D and E: Western blot analysis of kidney expression of corin and active b-catenin protein at days 4 and 7 after UUO. Representative Western blot analysis (D) and quantitative data (E) are presented. F: Linear regression graph of kidney corin and b-catenin protein expression at days 0, 4, and 7 after UUO. The Spearman correlation coefficient (R2) is shown. G: Representative micrographs using immunohistochemical staining for proprotein convertase subtilisin/kexin-6 (PCSK6) in kidney tissues at days 4 and 7 after UUO. Boxed area shows magnification of image. Arrows indicate PCSK6-positive tubular epithelial cells. Arrowheads in the enlarged boxed area indicate positive staining. H and I: Western blot analysis of PCSK6 protein in kidney at days 4 and 7 after UUO. Representative Western blot analysis (H) and quantitative data (I) are presented. Data are the means SEM (B, E, and I). n Z 6 animals per group (B, E, and I). *P < 0.05, **P < 0.01, and ***P < 0.001. Scale bars: 50 mm (C and G, main images); 200 mm (C and G, enlarged images).

Article Snippet: HKC-8 cells were treated with recombinant human transforming growth factor (TGF)-b1 ajp.amjpathol.org - The American Journal of Pathology Table 1 Used Antibodies in Experiments Antibodies Dilution for IHC/IF Dilution for WB Company Product code Anti-Wnt1 monoclonal antibody e 1:1000 Abcam (Cambridge, UK) ab15251 Anti-corin monoclonal antibody IHC: 1:200 1:1000 Abcam ab274323 Antieb-catenin nonphosphorylated (active) S37/T41 monoclonal antibody IHC: 1:2000 1:1000 Abcam ab246504 GAPDH monoclonal antibody e 1:5000 ProteinTech (Chicago, IL) 60004-1-Ig HA tag polyclonal antibody e 1:1000 ProteinTech 51064-2-AP MYC tag polyclonal antibody e 1:1000 ProteinTech 16286-1-AP DYKDDDDK tag recombinant antibody (binds to FLAG tag epitope) e 1:500 ProteinTech 80010-1-RR Smooth muscle actin polyclonal antibody IHC: 1:200 IF: 1:100 1:1000 ProteinTech 14395-1-AP Fibronectin monoclonal antibody IHC: 1:200 IF: 1:100 1:2000 ProteinTech 66042-1-Ig SNAIL1 polyclonal antibody e 1:500 ProteinTech 13099-1-AP PAI-1 polyclonal antibody e 1:1000 ProteinTech 13801-1-AP PCSK6 polyclonal antibody IHC: 1:100 1:500 Novus Biologicals (Centennial, CO) NBP1-05030 Collagen type I polyclonal antibody e 1:2000 ProteinTech 14695-1-AP CD31 IHC: 1:100 IF: 1:100 e Santa Cruz Biotechnology (Dallas, TX) sc-376764 PDGFRb IHC: 1:100 IF: 1:100 e Affinity Biosciences (China) AF6133 e, not applicable; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HA, hemagglutinin; IF, immunofluorescence; IHC, immunohistochemical; PAI-1, plasminogen activator inhibitor 1; PCSK6, proprotein convertase subtilisin/kexin-6; PDGFRb, platelet-derived growth factor receptor b; WB, Western blot. at 2 ng/mL for different periods of time, except as otherwise indicated in some experiments.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: iScience

Article Title: Semi-supervised analysis of myeloid and T cell behavior in ex vivo ovarian tumor slices reveals changes in cell motility after treatments

doi: 10.1016/j.isci.2023.106514

Figure Lengend Snippet: Key resources table

Article Snippet: Fibronectin Antibody (HFN7.1) [Alexa Fluor® 405] , Novus biologicals , Cat# NBP2-34633AF405; RRID: N/A.

Techniques: Control, Recombinant, Marker, Isolation, Blocking Assay, Plasmid Preparation, Sterility, Polymer, Conjugation Assay, Red Blood Cell Lysis, Software, Cell Culture