Journal: Scientific Reports

Article Title: Caldesmon-1–mediated actin dynamics is essential for osteogenic differentiation of aortic valve interstitial cells

doi: 10.1038/s41598-026-39938-x

Figure Lengend Snippet: Localisation of caldesmon-1 (CALD1) and α-smooth muscle actin (αSMA) proteins in normal and AS aortic valves. ( a ) Immunohistochemical staining with anti-CALD1 and anti-αSMA antibodies and histological analyses by haematoxylin and eosin (H&E) and Massons’ trichrome in normal and aortic valve stenosis (AS) valve leaflets. Scale bar = 100 μm. ( b ) Quantification of CALD1- and αSMA-positive areas in normal and AS valves ( n = 7). Statistical significance was determined using a Mann–Whitney U test (* P < 0.05). ( c ) Enlarged image of the CALD1 IHC shown in Fig. 1a. Calcified areas identified histologically are highlighted in red ( Left panel ). Immunofluorescence images of CALD1- (green) and αSMA- (red) double-positive VICs in AS valve (Non-calcified area and Calcified area) ( Right panel ). Scale bar = 50 μm.

Article Snippet: Membranes were incubated with primary antibodies, including anti-CALD1 (Proteintech, Catalogue no. 66693-I, 1:4000 dilution), anti-phosphoMLC (Cell Signaling Technology, Catalogue no. 3671, 1:5000 dilution), and anti-ACTB (Sigma-Aldrich, Catalogue no. A5441, 1:4000 dilution).

Techniques: Immunohistochemical staining, Staining, MANN-WHITNEY, Immunofluorescence

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 is a potential regulatory factor mediating Parkinson’s disease in mice: ( a ) Reproductive flowchart of BAP31 fl/fl and Slc6a3cre-BAP31 fl/fl mice. ( b ) GO annotation classification histogram in the substantia nigra pars compacta of BAP31 fl/fl and Slc6a3cre-BAP31 fl/fl mice. ( c ) The KEGG enrichment analysis of differentially expressed genes in the substantia nigra pars compacta of BAP31 fl/fl and Scl6a3 cre-BAP31 fl/fl mice. ( d ) The volcanic pattern analysis of these differentially expressed genes. ( e ) The Venn diagrams of the sample database alongside the two Parkinson’s disease databases, GSE8397 -DEGs and GSE20164 -DEGs.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques:

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 deficiency increased behavioral dysfunction in MPTP-treated mice: ( a ) mRNA expression of BAP31 in primary DA neurons from Slc6a3cre-BAP31 fl/fl and BAP31 fl/fl analyzed by real-time PCR (BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl , n = 3). ( b ) Protein expression of BAP31 in primary DA neurons from Slc6a3cre-BAP31 fl/fl and BAP31 fl/fl , detected by WB (BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl , n = 3). ( c – f ) Behavioral results of 6-month-old Slc6a3cre-BAP31 fl/fl and BAP31 fl/fl mice injected with saline or MPTP (half male and female). ( c ) Walking time of the balance beam test. ( d ) Pole climbing test. ( e ) Rotating rod experiment. ( f ) The open-field test. (Experimental group: BAP31 fl/fl mice, n = 12; Slc6a3cre-BAP31 fl/fl mice, n = 12; BAP31 fl/fl mice injected with MPTP, n = 12; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP, n = 12). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Injection, Saline

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 deficiency increased neuron loss induced by MPTP-treated mice: ( a ) Expression levels of NeuN and GFAP were detected by immunofluorescence assay. Scale bar = 50 μm. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP, n = 3. ( b ) Results of HE staining in the midbrain of mice. Scale bar = 500 or 100 μm. ( c ) Results of HE staining in the striatum of mice. Scale bar = 500 or 100 μm. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP, n = 3. ( d ) The ultrastructure of the nucleus in the midbrain and striatum of mice was observed by an electron microscope. Scale bar = 1 μm. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP n = 3. ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Expressing, Immunofluorescence, Injection, Staining, Microscopy

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 deficiency increased dopamine neuron loss in MPTP-treated mice: ( a , b ) TH content in the midbrain and striatum of mice was detected by immunohistochemistry. Scale bar = 500, 200, or 50 μm. ( c ) TH level in the midbrain and striatum of mice was detected by WB. ( d ) BAP31 deficiency affected the DAT level in the brain of MPTP-treated mice. Scale bar = 50 μm. ( e ) ELISA detected the DA, DOPAC, and HVA levels in the midbrain and striatum of the brains of mice. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Injection

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 deficiency results in MPTP-lesioned mitochondrial homeostasis in PD mice: ( a – e ) The ultrastructure of mitochondria in the midbrain and striatum of mice was observed using electron microscopy. Mitochondrial status was rated as I–IV, and the number of different grades was counted. Scale bar = 1 μm. ( f ) The content of BAP31, MFF, Drp1, FIS1, MFN1, and OPA1 in the midbrain and striatum was detected by WB. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Electron Microscopy, Injection

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 deficiency affected the expression of PINK1/Parkin pathway-related proteins in MPTP-treated mice: ( a ) GeneCard scores of PD-related genes, indicating 19 genes with scores above 90. ( b ) The mRNA levels of each protein in the mouse brain were analyzed by qPCR. ( c ) The protein expressions of BAP31, PINK1, and Parkin were detected and quantitatively analyzed by WB. ( d , e ) The results of immunofluorescence double staining indicated the fluorescence levels of PINK1 and Parkin in the midbrain and striatum of mice. Scale bar = 50 μm. Experimental groups: BAP31 fl/fl mice, n = 3; Slc6a3cre-BAP31 fl/fl mice, n = 3; BAP31 fl/fl mice injected with MPTP, n = 3; and Slc6a3cre-BAP31 fl/fl mice injected with MPTP, n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Expressing, Immunofluorescence, Double Staining, Fluorescence, Injection

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: BAP31 regulated the levels of PINK1 through EN1: ( a ) SH-SY5Y was transfected with siBAP31, and a Flag tag and WB were performed to observe changes in PINK1 protein levels. ( b ) Co-IP results indicate BAP31 and PINK1. ( c ) SH-SY5Y cells were transfected with siBAP31, and changes in SP1 and EN1 protein levels were observed by WB. ( d ) SH-SY5Y cells were transfected with Flag, and changes in SP1 and EN1 protein levels were detected by WB. ( e ) Correlation analysis of BAP31 and EN1. ( f ) WB analysis of PINK1 expression in cells transfected with siRNA-BAP31 and overexpressing EN1. ( g ) Analysis of the mRNA level of PINK1 expression in cells transfected with siRNA-BAP31 and those overexpressing EN1. ( h ) WB analysis of PINK1 expression in cells transfected with siRNA-EN1 and overexpressing BAP31. ( i ) Analysis of the mRNA level of PINK1 expression in cells transfected with siRNA-EN1 and overexpressing BAP31. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Transfection, FLAG-tag, Co-Immunoprecipitation Assay, Expressing

Journal: Cells

Article Title: BAP31 Modulates Mitochondrial Homeostasis Through PINK1/Parkin Pathway in MPTP Parkinsonism Mouse Models

doi: 10.3390/cells15020137

Figure Lengend Snippet: We used arrowheads (→) to indicate activation or positive regulation and T-shaped arrowheads (--|) to indicate inhibition or negative regulation. In this study, we found that BAP31 deficiency down-regulated the PINK1 transcription factor EN1, thereby impairing the PINK1–Parkin pathway. This disruption led to an imbalance in mitochondrial fission and fusion, mitochondrial dysfunction, loss of dopamine neurons, and ultimately exacerbated PD development.

Article Snippet: Antibodies to BAP31 (#12226-1-AP, 1:1000, Proteintech Group, Chicago, IL, USA), TH (#ab6211, 1:1000, Abcam, Cambridge, MA, USA), PINK1 (Santa Cruz;sc-517353, 1:1000, Signalway Antibody, College Park, MD, USA), BAP31 (11200-1-AP, 1:1000, Proteintech), PARK7 (11681-1-AP, 1:1000, Proteintech), PARK2 (14060-1-AP,1:1000, Proteintech), SP1 (WL02251, 1:1000, Wanlei Biotechnology, Shanghai, China), EN1 (ab108598, 1:1000, Abcam), and MFF (12186-1-AP, Proteintech) were used.

Techniques: Activation Assay, Inhibition, Disruption

Journal: Scientific Reports

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1038/s41598-025-18471-3

Figure Lengend Snippet: RO8191 induces embryo implantation in delayed implantation (DI) model mice. ( A ) The experimental procedure for the artificial DI mouse. Ovariectomy (OVX) and subcutaneous administration of siliconized medroxyprogesterone acetate (MPA) were performed on D3. A single injection of oil or RO8191 was performed on D7. Females were euthanized and dissected on D10. ( B ) Representative images of the gross uterine morphology. Blastocysts were recovered from oil-treated mice (white arrowheads). Implantation sites (yellow arrowheads on right panel) were visible in DI mice after RO8191 treatment. Scale bars: 1 cm for uterine images and 200 μm for blastocysts. ( C , D ) Implantation rate ( C ) and the number of implantation sites ( D ) in the oil and RO8191-treated DI mice. *Significantly different ( p < 0.0001). ( E ) Representative histological image of uterine cross-sections of implantation sites on D10 after RO8191 treatment in DI mice. Right panels are higher magnification of left panels indicated by yellow line. Lower panels indicated abnormal embryo development and decidual reaction. Scale bars: 100 μm. dec: decidua; em: embryo.

Article Snippet: The RO8191 ( T22142 , TargetMol, Boston, MA, USA, or SML1200, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals, or S3547, Sigma-Aldrich), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head) or sesame oil was performed at 1300 h on D7.

Techniques: Injection

Journal: Scientific Reports

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1038/s41598-025-18471-3

Figure Lengend Snippet: RO8191 induces decidual reaction in mice with genetic implantation failure. ( A ) The experimental procedure for the genetically modified mice. For control, a single oil injection was performed on D4. For cKOs, a single injection of oil or RO8191 was performed on D4. Females were euthanized and dissected on D7. ( B – D ) Uterine morphology of cKOs mice after the injection of oil or RO8191. The white arrowheads indicate the presumptive implantation sites, including uterine swelling. Scale bars: 1 cm. ( E ) The implantation rate in cKOs mice after oil or RO8191 administration. ( F ) The number of implantation sites in cKOs mice after the injection of oil or RO8191. ( G ) The size of implantation sites in cKOs mice after oil or RO8191 administration. *Significantly different ( p < 0.05).

Article Snippet: The RO8191 ( T22142 , TargetMol, Boston, MA, USA, or SML1200, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals, or S3547, Sigma-Aldrich), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head) or sesame oil was performed at 1300 h on D7.

Techniques: Genetically Modified, Control, Injection

Journal: Scientific Reports

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1038/s41598-025-18471-3

Figure Lengend Snippet: RO8191 activates STAT3, but not STAT1, in the endometrium. ( A ) Western blot analysis of STAT3 and phospho-STAT3 (p-STAT3) proteins after RO8191 treatment. The predicted band sizes are 89 kDa (STAT3α), 69 kDa (STAT3β), and 66 kDa (p-STAT3). ( B ) Western blot analysis of STAT1 and p-STAT1 after RO8191 treatment. A549 cells treated with IFN-γ were loaded as a control. The predicted band sizes are 91 kDa (STAT1α and p-STAT1α), 84 kDa (STAT1β and p-STAT1β). Blot images were obtained from one gel, and dividing lines indicate their separate origins. LE and ST protein lysates were pooled from at least three different individuals. ( C ) Representative immunohistochemical images of p-STAT3 expression in the uterus after RO8191 treatment. E2-treated uterus is shown as a control. Specimens were prepared at least three different individuals (two individuals in the case of E2). Scale bar: 100 μm. le: luminal epithelium; st: stroma.

Article Snippet: The RO8191 ( T22142 , TargetMol, Boston, MA, USA, or SML1200, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals, or S3547, Sigma-Aldrich), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head) or sesame oil was performed at 1300 h on D7.

Techniques: Western Blot, Control, Immunohistochemical staining, Expressing

Journal: Scientific Reports

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1038/s41598-025-18471-3

Figure Lengend Snippet: Effect of RO8191 on the STAT3 signaling pathway in knockout mice. ( A ) Histological characterization of uterine cross-sections from presumed implantation sites on D7. Images a’-d’ are higher magnifications of images a-d, with further magnification in the yellow frames in b’ and c’. Scale bars: 100 μm. dec: decidua; le: luminal epithelium; em: embryo; bv: blood vessels; hem: hemorrhage; li: leukocyte infiltration. ( B ) Representative images of alkaline phosphatase (ALPL) expression. Scale bar: 100 μm. em: embryo; le: luminal epithelium; st: stroma. ( C ) The experimental procedure for maintaining pregnancy in Lifr cKO mice. A single injection of RO8191 was performed on D4, and females underwent cesarean section (CS) on D20-22. A single injection of RO8191 was sufficient to establish pregnancy in Lifr cKO mice. Representative images show an Lifr cKO female on D15 and the offspring obtained by CS. ( D ) Representative images of p-STAT3 in Gp130 cKO and Lifr cKO uterus 6 h after RO8191 treatment (D4 1600 h). Scale bar: 100 μm. le: luminal epithelium; st: stroma.

Article Snippet: The RO8191 ( T22142 , TargetMol, Boston, MA, USA, or SML1200, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals, or S3547, Sigma-Aldrich), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head) or sesame oil was performed at 1300 h on D7.

Techniques: Knock-Out, Expressing, Injection