Journal: Scientific Reports

Article Title: Ovarian steroids modulate mRNA expression of ECM associated genes and collagen deposition induced by TGF β1 in equine endometrium in vitro

doi: 10.1038/s41598-024-84250-1

Figure Lengend Snippet: The effect of progesterone (P4; 10 − 7 M) or 17β-estradiol (E2; 10 − 9 M) on mRNA expression of collagen type 1 alpha chain 1 ( COL1A1) ( A ), collagen type 3 alpha chain 1 ( COL3A1) ( B ), matrix metalloproteases (MMP): MMP-1 ( C ), MMP-2 ( D ), MMP-3 ( E ), MMP-9 ( F ), and MMP-13 ( G ), and tissue inhibitors of matrix metalloproteases (TIMPs): TIMP-1 ( H ) and TIMP-2 ( I ) in equine endometrial fibroblasts. Data were analyzed by Student’s t-test and expressed as mean ± SEM. Asterisks designate statistical differences between treatments (* p < 0.05; ** p < 0.01; *** p < 0.001; **** P < 0.0001).

Article Snippet: The primary antibody against COL1 (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4ºC, and the secondary antibody horseradish peroxidase (HRP)-conjugated anti-rabbit (1:20,000; RRID: AB_2617138; P0448, DakoCytomation, Carpinteria, CA, USA) was incubated during 1.5 h at room temperature.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Ovarian steroids modulate mRNA expression of ECM associated genes and collagen deposition induced by TGF β1 in equine endometrium in vitro

doi: 10.1038/s41598-024-84250-1

Figure Lengend Snippet: Relative collagen type I alpha chain 2 ( COL1A2 ) mRNA expression of equine endometrial explants obtained from mares in the follicular phase (FP; n = 5) and treated for 24 ( A ) or 48 h ( B ) with medium alone (control), transforming growth factor β1 (TGF-β1; 10 ng/mL), TGF-β1 (10ng/mL) + 17β-estradiol (E2; 10 − 9 M) or E2 alone (10 − 9 M). Results were analyzed by one-way analysis of variance (ANOVA), followed by a Tukey’s multiple comparisons test and considered significant at p < 0.05 and shown as mean ± SEM. Asterisks represent significant differences relative to the respective control and asterisks above lines designate differences between treatments (* p < 0.05; ** p < 0.01; **** P < 0.0001).

Article Snippet: The primary antibody against COL1 (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4ºC, and the secondary antibody horseradish peroxidase (HRP)-conjugated anti-rabbit (1:20,000; RRID: AB_2617138; P0448, DakoCytomation, Carpinteria, CA, USA) was incubated during 1.5 h at room temperature.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: Ovarian steroids modulate mRNA expression of ECM associated genes and collagen deposition induced by TGF β1 in equine endometrium in vitro

doi: 10.1038/s41598-024-84250-1

Figure Lengend Snippet: Relative collagen type I (COL1) protein abundance in equine endometrial explants obtained from mares in the follicular phase (FP; n = 5) and treated for 24 ( A ) or 48 h ( B ) with medium alone (control), transforming growth factor β1 (TGF-β1; 10 ng/mL), TGF-β1 (10ng/mL) + 17β-estradiol (E2; 10 − 9 M) or E2 (10 − 9 M), respectively. Results were analyzed by one-way analysis of variance (ANOVA), followed by a Tukey’s multiple comparisons test and considered significant at p < 0.05 and shown as mean ± SEM. Asterisks represent significant differences relative to the respective control and asterisks above lines designate differences between treatments (* p < 0.05; ** p < 0.01; *** P < 0.001).

Article Snippet: The primary antibody against COL1 (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4ºC, and the secondary antibody horseradish peroxidase (HRP)-conjugated anti-rabbit (1:20,000; RRID: AB_2617138; P0448, DakoCytomation, Carpinteria, CA, USA) was incubated during 1.5 h at room temperature.

Techniques: Control

Journal: Scientific Reports

Article Title: Ovarian steroids modulate mRNA expression of ECM associated genes and collagen deposition induced by TGF β1 in equine endometrium in vitro

doi: 10.1038/s41598-024-84250-1

Figure Lengend Snippet: Relative collagen type I alpha chain 2 ( COL1A2 ) mRNA expression in equine endometrial explants obtained from mares in the mid-luteal phase (MLP; n = 5), and treated for 24 ( A ) or 48 h ( B ) with medium alone (control), transforming growth factor β1 (TGF-β1; 10 ng/mL), TGF-β1 (10ng/mL) + progesterone (P4; 10 − 7 M) or P4 alone (10 − 7 M). Results were analyzed by one-way analysis of variance (ANOVA), followed by a Tukey’s multiple comparisons test and considered significant at p < 0.05 and shown as mean ± SEM. Asterisks represent significant differences relative to the respective control and asterisks above lines designate differences between treatments (* p < 0.05; ** p < 0.01; *** P < 0.001).

Article Snippet: The primary antibody against COL1 (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4ºC, and the secondary antibody horseradish peroxidase (HRP)-conjugated anti-rabbit (1:20,000; RRID: AB_2617138; P0448, DakoCytomation, Carpinteria, CA, USA) was incubated during 1.5 h at room temperature.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: Ovarian steroids modulate mRNA expression of ECM associated genes and collagen deposition induced by TGF β1 in equine endometrium in vitro

doi: 10.1038/s41598-024-84250-1

Figure Lengend Snippet: Relative collagen type I (COL1) protein abundance in equine endometrial explants obtained from mares in the mid-luteal phase (MLP; n = 5) and treated for 24 ( A ) or 48 h ( B ) with medium alone (control), transforming growth factor β1 (TGF-β1; 10 ng/mL), TGF-β1 (10ng/mL) + progesterone (P4; 10 − 7 M) or P4 (10 − 7 M), respectively. Results were analyzed by one-way analysis of variance (ANOVA), followed by a Tukey’s multiple comparisons test and considered significant at p < 0.05 and shown as mean ± SEM. Asterisks represent significant differences relative to the respective control and asterisks above lines designate differences between treatments (* p < 0.05; *** P < 0.001).

Article Snippet: The primary antibody against COL1 (1:1000 diluted; RRID: AB_2891017, 20121, Novotec, Lyon, France) was incubated overnight at 4ºC, and the secondary antibody horseradish peroxidase (HRP)-conjugated anti-rabbit (1:20,000; RRID: AB_2617138; P0448, DakoCytomation, Carpinteria, CA, USA) was incubated during 1.5 h at room temperature.

Techniques: Control

Journal: bioRxiv

Article Title: Extracellular matrix guidance determines proteolytic and non-proteolytic cancer cell patterning

doi: 10.1101/2022.03.16.484647

Figure Lengend Snippet: Cell emigration patterns in the presence or absence of MMP inhibitor GM6001 after 48h culture. (A) Xz view of interface migration at indicated conditions and in 6 mg/ml collagen after 48h culture in the presence or absence of GM6001. Where present, the rim of the core is indicated by white dotted line. Graph, COL1 ¾ signal per cell (counted as 75 and 118 nuclei per condition from the invasion zone, but not the core; n=1). (B-G) Cell emigration parameters in the presence of GM6001 after 48h. (A) Gray dotted circles indicate size of depicted spheroid when imaged at 0h culture. (F) Area bordered by white line in top images indicates the area of the zoom-in in the respective lower image; orange arrowheads, interface; open arrowheads, cytoplasmic extensions; closed arrowheads, invaded nuclei. (C-E, G, H) Quantification of interface-guided and non-guided migration parameters as described in and . Data represent 1-4 measurements (dots/line) per condition per experiment (N=2). (I , J) Interface-mediated versus non-guided migration in the absence or presence of MMP activity. (I) Percentage of emigrated cells in the interface compared to total number of emigrated cells per spheroid. (J) Total numbers of interface-guided cells plotted against non-guided cells. Orange line indicates condition of MMP-independent guidance. (I, J) Data from bars depicted with “M” were derived from in silico cellular automata-based model. In vitro data were derived from experiments depicted in ; data represent 1-4 measurements (bar/square) per condition per experiment (N=2). (C-E, H) Solid line, median, (I , J) Bars and squares, mean; whiskers, SD. Mann-Whitney test with, when necessary, Holm-Sidak, ns: not significant; *: P-value < 0.05; **: P-value < 0.01. Scale bars: (A) 50 μm; (B, F) 250 μm.

Article Snippet: Affinity purified rabbit anti-collagen type I cleavage site antibody (Col1¾C short , Immunoglobe), Alexa-488-conjugated secondary pre-absorbed goat anti-rabbit IgG (Invitrogen), DAPI (Sigma) and Alexa-568-conjugated phalloidin (Invitrogen) were used.

Techniques: Migration, Activity Assay, Derivative Assay, In Silico, In Vitro, MANN-WHITNEY