lc ms ms  (R&D Systems)

Journal: Nature Communications

Article Title: Androgens drive SLC1A5-dependent metabolic reprogramming in polycystic ovary syndrome

doi: 10.1038/s41467-025-62951-z

Figure Lengend Snippet: a Schematic diagram of the DHEA-induced PCOS-like model. Created in BioRender. Ye, L. (2025) https://BioRender.com/yb2yu8f . b–d mRNA expression of Slc1a5 ( b ), Hdac5 ( c ), and Cyp19a1 ( d ) in ovaries collected from Control and DHEA-treated mice determined by qPCR ( n = 12 for each group). e – h Western blots ( e ) and densitometric analysis of SLC1A5 ( f ), HDAC5 ( g ), and CYP19A1 ( h ) expression relative to α-Tubulin in the ovaries of Control and DHEA-treated mice ( n = 5 for each group). i Schematic diagram of the PAMH model. Created in BioRender. Ye, L. (2025) https://BioRender.com/0wojt74 . j Levels of glutamine in the ovaries of F1 female offspring of PBS and AMH-treated ( n = 5 for each group) dams. k – m mRNA expression of Slc1a5 ( k ) Hdac5 ( l ) and Cyp19a1 ( m ) in the ovaries of F1 female offspring of PBS and AMH-treated dams determined by qPCR ( n = 15 for each group). n – q Western blots ( n ) and densitometric analysis of SLC1A5 ( o ) HDAC5 ( p ) and CYP19A1 ( q ) expression relative to β-Actin in the ovaries of PBS and AMH-treated mice ( n = 5 for each group). For statistical analysis, Mann-Whitney tests were performed in ( b, c, m ); and Student’s t tests were performed in ( d, f –h, j–l, o–q ). All tests were two-sided. The biological replicates are used for statistical analysis in ( b –h, j–q ). Source data are provided as a Source Data file.

Article Snippet: The control group received intraperitoneal injections of PBS (10 μL/g body weight), while the PAMH group was administered AMH (0.12 mg/kg body weight, R&D Systems, 1737-MS-010, MN, USA) via intraperitoneal injection.

Techniques: Expressing, Control, Western Blot, MANN-WHITNEY

Journal: Nature Communications

Article Title: Gene therapy with feline anti-Müllerian hormone analogs disrupts folliculogenesis and induces pregnancy loss in female domestic cats

doi: 10.1038/s41467-025-56924-5

Figure Lengend Snippet: a , b The cleavage recognition motif (RAQR) in feline AMH was modified to the more efficient RKKR sequence. The G561S gain-of-function point mutation was then incorporated into the fAMH_RKKR construct to generate fAMH_RKKR/G561S. ( c ) To determine the effects on fAMH synthesis, conditioned medium from HEK293T cells transfected with either wild-type or mutant constructs was analyzed by reducing Western blot. Blots were probed with mAb-5/6, targeted to mature AMH. Recombinant human AMH was used as a positive control. Western blot shown in ( c ) is representative of three separate transfection experiments. ( d ) SMAD1/5/9-dependant luciferase reporter (BRE-luc) activity in AMH-responsive HEK293T cells following treatment with diluted conditioned medium containing fAMH variants. Luciferase activity is presented as the mean ± SD of quadruplicates ( n = 4) from a representative experiment, relative to an adjusted value of 1.0 for the mean of wells which received fresh medium alone. The experiment was repeated 4 times. Data were analyzed using one-way ANOVA with Tukey’s post hoc test (GraphPad Prism v.10). P = 0.0146 (*), P = 0.0015 (**) and P = 0.0009 (***). Source data are provided as a Source Data file.

Article Snippet: Recombinant human AMH mature protein (R&D Systems, Minneapolis, MN, United States) was used as a positive control and as standards of known mass.

Techniques: Modification, Sequencing, Mutagenesis, Construct, Transfection, Western Blot, Recombinant, Positive Control, Luciferase, Activity Assay

Journal: Nature Communications

Article Title: Gene therapy with feline anti-Müllerian hormone analogs disrupts folliculogenesis and induces pregnancy loss in female domestic cats

doi: 10.1038/s41467-025-56924-5

Figure Lengend Snippet: a Based on ELISA measurements, ~1 μl of serum collected at various times (day 0, day 42, day 126 and day 239) from cat #985, in the fAMH_RKKR/G561S group, was diluted and analyzed by Western blotting, with samples run under reducing conditions. Serum collected on days 0 and 239 from cat #134, in the control group, was run as a negative control. Recombinant mature human AMH was used as a positive control. Blot was probed with mAb-5/6, targeted to the AMH mature domain. b Serum from cat #985 in the fAMH_RKKR/G561S group was collected 2 weeks prior (day 0) and at different times (4- (day 42), 16- (day 126) or 32- (day 239) weeks) after AAV delivery. Based on AMH ELISA measurements, serum was diluted 1:100 in fresh medium and used to treat HEK293T cells transfected with a SMAD1/5/9-responsive transcriptional reporter (BRE-luc) and AMH receptors. Luciferase activity for cells treated with diluted cat serum is presented as the mean ± SD of triplicates ( n = 3) from a representative experiment, relative to an adjusted value of 1.0 for the mean of the wells which received medium alone ( n = 6). The experiment was repeated 4 times. Data were analyzed using one-way ANOVA with Tukey’s post hoc test (GraphPad Prism v.10). P < 0.0001 (****). Source data are provided as a Source Data file.

Article Snippet: Recombinant human AMH mature protein (R&D Systems, Minneapolis, MN, United States) was used as a positive control and as standards of known mass.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Control, Negative Control, Recombinant, Positive Control, Transfection, Luciferase, Activity Assay