Journal: REPRODUCTION

Article Title: Involvement of VCAM1 in the bovine conceptus adhesion to the uterine endometrium

doi: 10.1530/rep-13-0655

Figure Lengend Snippet: Figure 5 Expression of VCAM1 receptor, integrin a 4 (ITGA4), by bovine conceptuses. (A) Changes in ITGA4 mRNAs in days 17, 20, and 22 conceptuses (P17, P20, and P22 respectively). Note that ITGA4 transcripts were found minimal at the uterine epithelium and stroma. Values represent meanGS.E.M. from three independent samples with duplicates within a day of conceptus collection. **Statistically significant differences in mRNA levels (P!0.01). (B) Immunohisto- chemical analysis of ITGA4 (a, b, c, and d) or VCAM1 (e, f, g, and h) expression in the bovine uterus obtained from day 22 pregnant animals. Tissue sections (10 mm) from day 22 uteri were immunostained for ITGA4 using an anti-ITGA4 antibody (a and c) or normal mouse IgG as a negative control at low (b) and higher magnification (d). (c) Observation of boxed area in B-a at a higher magnification. Tissue sections were immunostained for VCAM1 using an anti-VCAM1 antibody (e and g), or normal rabbit IgG as a negative control at low (f) and higher magnification (h). (g) Observation of boxed area in B-e at a higher magnification. Epi, endometrial luminal epithelium; St, endometrial stroma; Tr, trophoblast. Black scale barZ200 mm and white scale barZ40 mm.

Article Snippet: After 30 min of incubation with 10% normal goat serum, the sections were incubated at 4 8C overnight with a rabbit anti-human VCAM1 polyclonal antibody (1:100 dilution, 0.5 mg/ml, ab106777, Abcam, Cambridge, MA, USA), a mouse anti-human CD49d (ITGA4) antibody (1:100 dilution, 1 mg/ml, MCA697GA, AbD Serotec, Hercules, CA, USA), or normal mouse IgG (1:40 dilution, 0.4 mg/ml, sc-2025, Santa Cruz Biotechnology, Inc.) or normal rabbit IgG (1:80 dilution, 0.4 mg/ml, sc-2027, Santa Cruz Biotechnology, Inc.) as a Reproduction (2014) 148 119–127 A negative control.

Techniques: Expressing, Negative Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Pneumolysin-induced lung injury is independent of leukocyte trafficking into the alveolar space.

doi: 10.4049/jimmunol.173.2.1307

Figure Lengend Snippet: FIGURE 4. Effect of adhesion-blocking Abs or transient neutropenia on PLY-induced neutrophilic alveolitis and lung permeability increase in in- tact mice. A, Mice were either left untreated (0-h time point) or treated with PLY in the absence of function-blocking Abs (40 ng/mouse; f) or received i.v. injections of function-blocking anti-CD18 Abs () or anti-CD18 plus anti-CD49d Abs (v) or were pretreated with anti-Gr-1 to induce neutro- penia (z) followed by intratracheal application of PLY (40 ng/mouse). Mice were sacrificed at the indicated time points and subjected to BAL for determination of BAL fluid leukocyte differentials. B, Mice were either treated with PLY alone (f in B) or were pretreated with anti-CD18 and anti-CD49d Abs (v in B) or were made transiently neutropenic (z in B) followed by PLY application. One hour before sacrifice, mice then re- ceived FITC-labeled albumin i.v. to determine lung vascular leakage. , p 0.01 vs PLY-alone treatment. Values are given as mean SD of five experiments. AU, Arbitrary unit.

Article Snippet: Rat anti-mouse CD49d (VLA-4; clone PS/2; isotype IgG2b) was purchased from Serotec (Munich, Germany).

Techniques: Blocking Assay, Permeability, Labeling

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ITGA4 , Proteintech , Cat#19676-1-AP; RRID: AB_10640907.

Techniques: Recombinant, SYBR Green Assay, Protease Inhibitor, Stripping Membranes, cDNA Synthesis, Chromatin Immunoprecipitation, Bicinchoninic Acid Protein Assay, Plasmid Preparation, Purification, Expressing, Software

Journal: Life sciences

Article Title: Collagen matricryptin promotes cardiac function by mediating scar formation.

doi: 10.1016/j.lfs.2023.121598

Figure Lengend Snippet: Fig. 4. p1159 promotes fibroblast migration and this may occur via integrin-α4 (Itgα4). A. LV fibroblasts treated with p1159 (100 nM in serum free media, SFM) presented a significant increase in migration rate, which was similar to the positive control (10 % FBS), demonstrating strong pro-migratory properties. *p < 0.05 p1159 100 nM and 500 nM versus negative control (SFM); n = 6/group. B. p1159 in vivo treatment induces overexpression of Itgα4 post-MI. Left panel: mRNA levels n = 6/group; right panel: representative immunoblot image, TP = total protein stain (loading control), n = 4/group. **p < 0.01. C. The neutralizing antibody against Itga4 inhibited p1159 induced fibroblast migration. Positive control (10 % FBS), negative control (SFM), and p1159 (100 nM and 500 nM) were treated ± Itgα4 neutralizing antibody (Itga4i). The impedance values were normalized before incubation with Itga4i and after wound. *p < 0.05 represents when both concentrations of p1159 statistically differed from its respective groups with Itga4i, and from the negative controls; n = 6/group. D. Cardiac fibroblasts stimulated with p1159 overexpress Itga4 mRNA and this effect is abolished in the presence of Itga4i. Left panel: mRNA levels n = 6/group; right panel: representative immunoblot image, TP = total protein stain (loading control), n = 4/group. *p < 0.05 E. Itgα4 (red) co-localized with p1159 (green, FITC labeled) in human cardiac fibroblasts (HCF) and that effect was inhibited in the presence of Itga4i. Blue = nuclei, yellow = co-localization. Scale bar = 5 μm. All graphs display average ± SEM. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Cell migration was recorded in real time for 48 h. For the experiments testing integrin alpha 4 (Itgα4) as the receptor for p1159, cardiac fibroblasts were cultured in SFM with Itgα4 blocking antibody (#NBP1-26661, Novus Biologicals; Itgα4i) 15 μg/mL for 2 h before wound induction, followed by treatment as listed in Table 1.

Techniques: Migration, Positive Control, Negative Control, In Vivo, Over Expression, Western Blot, Staining, Control, Incubation, Labeling

Journal: Life sciences

Article Title: Collagen matricryptin promotes cardiac function by mediating scar formation.

doi: 10.1016/j.lfs.2023.121598

Figure Lengend Snippet: Fig. 5. p1159-induced migration may occur via Rho GTPase pathways. A. p1159-treatment reduced RhoGDI expression, and this effect was dependent on Itgα4. Gene expression of cardiac fibroblasts treated with p1159 100 nM ± Itgα4 blocking antibody (Ab) and negative control (SFM). *p < 0.05 versus negative control (SFM), #p < 0.05 versus 100 nM p1158/59. B. p1159 induces RhoA activation in fibroblasts. Representative immunofluorescence images of fibroblasts treated with negative control (SFM), positive control (10 % FBS) and SFM ± p1159 100 nM or 500 nM, indicating stimulation of Itgα4 receptor (green) and activation of RhoA (red). Right panels: Representative images of the same groups incubated 2 h prior with blocking antibody against Itgα4 (Itga4 Ab), indicating that both Itgα4 receptor and RhoA signals were strongly decreased by fibroblasts treated with p1159 in the presence of the Ab. Magnification/scale bars: 20× = 50 μm; 40× = 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Cell migration was recorded in real time for 48 h. For the experiments testing integrin alpha 4 (Itgα4) as the receptor for p1159, cardiac fibroblasts were cultured in SFM with Itgα4 blocking antibody (#NBP1-26661, Novus Biologicals; Itgα4i) 15 μg/mL for 2 h before wound induction, followed by treatment as listed in Table 1.

Techniques: Migration, Expressing, Gene Expression, Blocking Assay, Negative Control, Activation Assay, Immunofluorescence, Positive Control, Incubation

Journal: eLife

Article Title: Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

doi: 10.7554/eLife.60933

Figure Lengend Snippet:

Article Snippet: Antibody , CD49d (α4) (mouse monoclonal) , Fluidigm , Cat#3141004B , (1 μg/100 μL).

Techniques:

Journal: eLife

Article Title: Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

doi: 10.7554/eLife.60933

Figure Lengend Snippet:

Article Snippet: Antibody , CD49d (α4) (mouse monoclonal) , Fluidigm , Cat#3141004B , (1 μg/100 μL).

Techniques: