Journal: Frontiers in Microbiology

Article Title: Morpho-Phylo Taxonomy of Novel Dothideomycetous Fungi Associated With Dead Woody Twigs in Yunnan Province, China

doi: 10.3389/fmicb.2021.654683

Figure Lengend Snippet: Taxa used in the phylogenetic analysis of Teichosporaceae and their corresponding GenBank numbers.

Article Snippet: Misturatosphaeria aurantonotata , ATCC 42522 , NA , U43479 , U43461 , AY485625 , NA.

Techniques:

Journal: Cardiovascular Research

Article Title: Doxorubicin-induced cardiovascular toxicity: a longitudinal evaluation of functional and molecular markers

doi: 10.1093/cvr/cvad136

Figure Lengend Snippet: Proteomic analysis of aortic samples from DOX-treated mice at Weeks 2 and 6 and validation of two candidate biomarkers with western blotting. Plots of differentially expressed proteins at Weeks 2 and 6 ( A ). For A , horizontal grey line indicates significance threshold ( P < 0.05); up- and down-regulated proteins are indicated in blue and red, respectively; differentially expressed proteins are abbreviated (full names are listed in abbreviations list). Western blotting shows and corroborates higher levels of SERPINA3 in DOX-treated mice at Weeks 2 and 6, which persist at Week 9 ( B ). Western blotting also confirms higher levels of THBS1 in the DOX-treated group at Weeks 2 and 6 and shows a trend ( P = 0.052) towards increased THBS1 levels at Week 9 ( B ). Representative western blot images for SERPINA3 and THBS1 at Weeks 2, 6, and 9 ( C ). For C , ‘V’ and ‘D’ indicate lanes with vehicle and DOX aortic samples, respectively. D* on the blot at Week 9 contains a DOX sample for which no protein was detected and was excluded from analysis as such. For A , n = 8 in vehicle and DOX group for both Week 2 and Week 6. For B , vehicle group: n = 5, n = 6, and n = 6 at Weeks 2, 6, and 9, respectively; DOX group: n = 5, n = 6, and n = 5 at Weeks 2, 6, and 9, respectively. For B , Mann–Whitney U test for each time point. * and ** P < 0.05 and 0.01. Abbreviations for differentially expressed proteins in proteomics: SERPINA1E, α-1-antitrypsin; HMGN2, Non-histone chromosomal protein HMG-17; SERPINA3N, α-1-antichymotrypsin orthologue n; MT2, Metallothionein-2; MBL2, Mannose-binding protein C; MT1, Metallothionein-1; THBS1, Thrombospondin-1; FTL1, Ferritin light chain 1; COL14A1, Collagen α-1 (XIV) chain; LGALS3BP, Galectin-3-binding protein; RPL26, 60S ribosomal protein L26; IGHM, Immunoglobulin heavy constant µ; HAMP, Hepcidin; APOA2, Apolipoprotein A-II; HBA, Hemoglobin subunit α; HBB-B1, Hemoglobin subunit β-1; CA1, Carbonic anhydrase 1; KRT15, Keratin (type I cytoskeletal 15); SPTA1, Spectrin α; CA2; Carbonic anhydrase 2; RPL6, 60S ribosomal protein L6; VWA1, von Willebrand factor A domain-containing protein 1; RPL37A, 60S ribosomal protein L37a; CAPG, Macrophage-capping protein; RAD23A, UV excision repair protein RAD23 homolog A; IGHG1, Ig γ-1 chain C region (membrane-bound form); SLC4A1, Band 3 anion transport protein; ACLY, ATP-citrate synthase; ATPA1, Sodium/potassium-transporting ATPase subunit alpha-1.

Article Snippet: For validation of anti-THBS1 and anti-SERPINA3 primary antibodies, recombinant mouse THBS1 (catalogue number 7859-TH-050) and SERPINA3 (catalogue number 4709-PI-010) proteins were used as positive controls, which were purchased from Bio-Techne (Dublin, Ireland).

Techniques: Western Blot, MANN-WHITNEY, Binding Assay, Membrane

Journal: Cardiovascular Research

Article Title: Doxorubicin-induced cardiovascular toxicity: a longitudinal evaluation of functional and molecular markers

doi: 10.1093/cvr/cvad136

Figure Lengend Snippet: Quantification of SERPINA3 and THBS1 levels in plasma of control and AICT patients using ELISA. Schematic overview of the patient cohorts ( A ). AICT patients exhibited higher SERPINA3 and THBS1 levels compared with the control group ( B ). Both SERPINA3 and THBS1 showed an inverse Pearson correlation with LVEF ( C ). For B , data are presented as Tukey box plots with median (horizontal line) and the 25th and 75th percentile whiskers. For C , white and red dots represent control and AICT patients, respectively. n numbers are shown in A . For B , Mann–Whitney U test. * and **** P < 0.05 and 0.0001.

Article Snippet: For validation of anti-THBS1 and anti-SERPINA3 primary antibodies, recombinant mouse THBS1 (catalogue number 7859-TH-050) and SERPINA3 (catalogue number 4709-PI-010) proteins were used as positive controls, which were purchased from Bio-Techne (Dublin, Ireland).

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: Thrombospondin production by the ovulatory follicle. Granulosa cells were aspirated from monkey ovarian follicles after ovarian stimulation before (0), 12, 24, or 36 h after administration of an ovulatory dose of hCG; additional monkeys received hCG and the PTGS2 inhibitor celecoxib (36+C) 36 h before follicle aspiration. Granulosa cells were assessed by qPCR for mRNA levels of THBS1 (A) , THBS2 (B) , and THBS4 (C) . All THBS mRNA levels are expressed relative to BACT . Granulosa cell lysates were assessed for THBS1 (D,E) and THBS4 (D,F) by western blotting and are expressed relative to pan-actin (D) . For (A–C,E,F) , data are expressed as mean + SEM, n = 3–7 samples/treatment. Within each panel, groups with no common letters are different by ANOVA and Duncans post hoc test, p < 0.05.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: Western Blot

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: THBS1 and THBS4 immunodetection in monkey ovarian follicles. Immunocytochemical detection (brown) of THBS1 (A–G) and THBS4 (H–N) was localized to the granulosa cell layer of ovulatory follicles obtained before (0; A,H) and 12 (B,I) , 24 (C,J) , and 36 (D,K) hours (h) after hCG administration as well as 36 h after administration of hCG and celecoxib (36+C h; E,L ). Images shown are representative of n-3-6 ovaries/treatment. Granulosa cell immunodetection of THBS1 (F) and THBS4 (M) was reduced after preabsorption of the primary antibody with recombinant human THBS1 or THBS4 and similar to staining observed with no primary antibody (G,N) . Nuclei are counterstained blue. All panels are oriented as shown in (A) , with stroma (st) in lower left, granulosa cell (gc, arrow) layer central, and follicle antrum (an) in upper right. Arrowheads indicate stromal staining with antibody against THBS1 (A,E) , THBS4 (J) , and no primary antibody (G,N) . Images in (A–N) are at the same magnification; bar in (A) is 50 μm. Lower magnification images shows immunocytochemical detection of THBS1 (O) and no primary antibody (P) stained sections of an ovary obtained after ovarian stimulation and 36 h hCG. Staining is apparent in stromal vessels ( O , arrows). Non-specific staining is indicated near the granulosa cell basement membrane and in vessel lumens in (O,P) (arrowheads); bar in (A) is 100 μm for images in (O,P) .

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: Immunodetection, Recombinant, Staining, Membrane

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: THBS1 is pro-angiogenic in vitro . Monkey ovarian microvascular endothelial cells (mOMECs) were treated with human THBS1 protein at concentrations of 0.001–1 nM or no THBS1 (0 nM) as indicated. (A–C) Migration was assessed 24 h after plating on a porous membrane and treatment with THBS1. mOMECs (arrow) which migrated through pores (arrowhead) were stained with hematoxylin and eosin, photographed, and counted. Representative membranes from 0 nM (B) and 1 nM (C) treatment groups are shown. (D–F) Proliferation was assessed by Ki67 immunodetection in mOMECs cultured for 24 h with THBS1. Ki67 positive (arrows) and negative (arrowhead) cells are indicated in representative images from mOMECs treated with 0 nM (E) and 1 nM (F) THBS1. Data are expressed as a percentage of Ki67 positive cells among all cells counted. (G–M) Sprout formation in response to THBS1 treatment was determined after 1 day (G,I) and 2 days (H,J,L,M) in vitro . mOMECs coating a polymer bead in vitro before THBS1 treatment (Day 0; K ) shows absence of sprouts. Arrows indicate representative sprouts on Day 2 of treatment with no THBS1 (L) and 0.1 nM THBS1 (M) . Images were quantified for both the number of sprouts (sprouts/bead; G,H ) and sprout length in μm (H,J) . For (A,D,G–J) , data are expressed as mean + SEM, n = 3–5 samples/treatment. Within each panel, groups with no common letters are different by ANOVA and Duncans post hoc test, p < 0.05.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: In Vitro, Migration, Membrane, Staining, Immunodetection, Cell Culture, Polymer

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: Confirmatory experiments for the intrafollicular antibody injections. Serum levels of estradiol (A) and progesterone (B) were not different between animals receiving intrafollicular injection of the THBS1 antibody and animals receiving control IgG. For each day, hormone levels were compared by unpaired t -test. Data are expressed as mean + SEM, n = 4/group. (C) THBS1 antibody reduces THBS1 (1 nM)-stimulated mOMEC migration in vitro . Within each panel, groups with no common letters are different by ANOVA with 1 repeated measure and Duncans post hoc test, p < 0.05. Data are expressed as mean + SEM, n = 3/group.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: Injection, Control, Migration, In Vitro

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: Follicle rupture and oocyte release are compromised after intrafollicular injection with an antibody against THBS1. Ovarian surface at the time of ovary removal after intrafollicular injection of control IgG (A) or THBS1 antibody (B–D) . Oocytes were located within a THBS1 antibody-injected follicle (E) and on the surface of a THBS1 antibody-injected follicle (F) . Rupture site after intrafollicular injection of control IgG (G) . Rupture sites in follicles injected with the THBS1 antibody were either small (H) or absent (I) . Tissues shown in (E–I) were stained with hematoxylin and eosin. Oocytes (E,F) shown at approximately maximal diameter. Images in (G–I) are at the same magnification; bar in (I) = 0.5 mm. (J) Rupture site area after intrafollicular injection with control IgG (IgG) or THBS1 antibody. Groups are different by two-tailed unpaired t -test as indicated by * p < 0.05. Data are expressed as mean + SEM, n = 4/group.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: Injection, Control, Staining, Two Tailed Test

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: Oocyte retention and follicle rupture.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Thrombospondin 1 (THBS1) Promotes Follicular Angiogenesis, Luteinization, and Ovulation in Primates

doi: 10.3389/fendo.2019.00727

Figure Lengend Snippet: Angiogenesis and luteinization are compromised after intrafollicular injection with an antibody against THBS1. Histological sections of ovaries shown in were immunostained for VWF (brown) to assess angiogenesis; hematoxylin counterstain. (A,B) Endothelial cell invasion into the granulosa cell layer of a control IgG-injected follicle (A) and THBS1 antibody-injected follicle (B) shows VWF+ cells within the granulosa cell (gc) layer. Arrows indicate VWF+ cells which have invaded furthest from the stroma into the granulosa cell layer. Single arrowheads indicate stromal vessels. Double arrowheads indicate capillary luminal spaces with red blood cells in the control-IgG injected follicle only. (C) shows representative measurements of granulosa cell layer thickness (brown lines) and endothelial cell invasion (green lines). (A–C) are oriented as shown in (A) , with stroma (st) at bottom, granulosa cell (gc) layer central, and follicle antrum (an) at top. Images in (A–C) are at the same magnification; bar in (B) is 100 μm. Granulosa cell layer thickness (D) , endothelial cell invasion (E) , and the percent of the granulosa cell layer penetrated by endothelial cells (F) after intrafollicular injection with control IgG (IgG) or THBS1 antibody. Groups are different by two-tailed unpaired t -test as indicated by * p < 0.05. Data are expressed as mean + SEM, n = 4/group. (H,J) 3D modeling of endothelial cells (white on black background) is shown alongside (G,I) representative VWF immunostained ovarian sections after intrafollicular injection with control IgG (G,H) or THBS1 antibody (I,J) . Green arrows indicate stromal vessels, green arrowheads indicate capillary-like structures that connect to a stromal vessel, and yellow arrowheads indicate endothelial cells that lack connect to a stromal vessel. (G–J) are oriented with antrum at the top, granulosa cells central, and stroma at the bottom of each image/model; (G,I) at the same magnification.

Article Snippet: Proteins were transferred to a polyvinylidene fluoride membrane (Immobilon; Millipore, Billerica, MA) and probed using antibodies against THBS1 (0.01 μg/ml; goat; R&D Systems), THBS4 (0.01 μg/ml; goat; R&D Systems), or actin (5 μg/ml; mouse; Millipore) ( ).

Techniques: Injection, Control, Two Tailed Test