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: Advanced Healthcare Materials

Article Title: Efficacy of pH‐Responsive Surface Functionalized Titanium Screws in Treating Implant‐associated S. aureus Osteomyelitis with Biofilms Formation

doi: 10.1002/adhm.202403261

Figure Lengend Snippet: In vitro and in vivo biocompatibility and osseointegration evaluation of the functionalized titanium screws. A,B) Representative western blot images of the expression of osteogenesis‐related proteins and C,D) corresponding quantitative analyses ( n = 3). E) Immunofluorescence images of RUNX2 and COL1 staining of MEFs in different groups. F) Schematic diagram of the osseointegration and biocompatibility test of functionalized screws in an uninfected bone defect model. G) The implantation process of functionalized screws in rabbit tibial plateau. H) The micro‐CT 3D reconstructed images and I) bone tissue volume/total tissue volume (BV/TV) analysis from micro‐CT results of the newly formed bone surrounding the screws after 56 days of implantation ( n = 3). J) The methylene blue‐basic magenta staining and corresponding K) bone‐implant contact (BIC) ratio analysis of the non‐decalcified bone sections for osseointegration assay ( n = 3). Green arrows indicate gaps between the screw and bone tissue. Blue arrows indicate bone‐implant contact areas. In I,J), The data are shown as the means ± SDs, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Additionally, the MEFs treated as described above were fixed with 4% polyformaldehyde for 15 min and permeabilized with 0.1% Triton‐X for 15 min. After incubated with primary antibody RUNX2 (Abmart, dilution 1:500) or COL1 (Abmart, dilution 1:500) at 4 °C overnight, the MEFs were exposed to secondary antibody (Servicebio, dilution 1:500) for one hour at room temperature.

Techniques: In Vitro, In Vivo, Western Blot, Expressing, Immunofluorescence, Staining, Micro-CT

Journal: Advanced Science

Article Title: WAC Facilitates Mitophagy‐mediated MSC Osteogenesis and New Bone Formation via Protecting PINK1 from Ubiquitination‐Dependent Degradation

doi: 10.1002/advs.202404107

Figure Lengend Snippet: WAC was up‐regulated during the osteogenic differentiation of MSCs. a) ARS Staining and quantification throughout MSC osteogenic differentiation; b) ALP Staining and quantification during the progression of MSC osteogenesis; c) Immunofluorescence Staining for COL1 (red) during osteogenic induction. Quantification is presented in the right panel (Scale bar = 50 µm); d) Relative mRNA levels of WAC assessed by qRT‐PCR at different time points during MSC osteogenesis; e) Protein levels of WAC and osteogenic markers (RUNX2, Osterix, OCN) during MSC osteogenic differentiation; f) Pearson correlation analysis depicting the relationship between WAC expression and quantification of RUNX2, Osterix, and OCN levels during MSC osteogenic differentiation; g) qRT–PCR and Western blotting to detect WAC mRNA and protein levels in bone marrow MSCs from nonosteoporotic patients and patients with osteoporosis; h) HE staining and immunohistochemical staining for WAC in the femurs of SAMR1 mice and SAMP6 mice (Scale bar = 100 µm). All data are presented as the means ± SD, n = 6 per group in (a, b, c), n = 5 in (g), n = 9 in (d, e, g), n = 12 in (f). Statistical differences were determined using Student's t ‐test or ANOVA. ** p < 0.01 and *** p < 0.001.

Article Snippet: Subsequently, the cells were treated with Triton X‐100 for 15 min at room temperature and incubated with goat serum for 30 min. Primary antibodies against COL1 (Cell Signaling Technology, Cat. No. 66 948), LC3B (Abcam, Cat. No. Ab192890), or TOM20 (Abcam, Cat. No. Ab186735) were added and left to incubate overnight at 4 °C.

Techniques: Staining, Immunofluorescence, Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemical staining

Journal: Advanced Science

Article Title: WAC Facilitates Mitophagy‐mediated MSC Osteogenesis and New Bone Formation via Protecting PINK1 from Ubiquitination‐Dependent Degradation

doi: 10.1002/advs.202404107

Figure Lengend Snippet: WAC positively regulated the osteogenic differentiation of MSCs in vitro. WAC is modulated in MSCs through SiRNA and Lentivirus. a) MSCs were cultured in osteogenic medium after transfection with SiRNA. ARS staining, ALP staining, and quantification were performed on day 12; b) Western blotting for protein levels of osteogenesis‐related markers (RUNX2, Osterix, OCN). Quantification is presented in the right panel; c) Immunofluorescence staining for COL1 (red) after SiRNA transfection. Quantification is shown in the right panel (Scale bar = 50 µm); d) Overexpression lentivirus of WAC transfected into MSCs, followed by culture in osteogenic medium. ARS staining, ALP staining, and quantification conducted on day 12; e) Western blotting for protein levels of osteogenesis‐related markers after overexpression lentivirus transfection; f) Immunofluorescence staining for COL1 (red) after overexpression lentivirus transfection. Quantification is shown in the right panel (Scale bar = 50 µm). All data are presented as the means ± SD, n = 6 per group in (a, c, d, f), n = 9 per group in (b, e). Statistical differences were determined using Student's t ‐test or ANOVA. ns not statistically significant, ** p < 0.01 and *** p < 0.001.

Article Snippet: Subsequently, the cells were treated with Triton X‐100 for 15 min at room temperature and incubated with goat serum for 30 min. Primary antibodies against COL1 (Cell Signaling Technology, Cat. No. 66 948), LC3B (Abcam, Cat. No. Ab192890), or TOM20 (Abcam, Cat. No. Ab186735) were added and left to incubate overnight at 4 °C.

Techniques: In Vitro, Cell Culture, Transfection, Staining, Western Blot, Immunofluorescence, Over Expression