Journal: The Journal of Reproduction and Development

Article Title: Structure and functions of the placenta in common minke ( Balaenoptera acutorostrata ), Bryde’s ( B. brydei ) and sei ( B. borealis ) whales

doi: 10.1262/jrd.2015-005

Figure Lengend Snippet: Immunostaining of P450arom in the sei whale placenta (L 144.2 cm). A: Areolar part of chorionic villi. B: Enlargement of the fetomaternal attachment area. P450arom immunoreactivity was detected in trophoblast cells, but not in areolar trophoblast cells. a: Areola. c: Chorionic villi. e: Endometrium. Arrows indicate trophoblast cells. Arrowheads indicate areolar trophoblast cells. * Transitional part between trophoblast cells and areolar trophoblast cells. Bar = 100 μm (A), 10 μm (B).

Article Snippet: Sections were incubated overnight with the polyclonal anti-rat cytochrome P450 side chain cleavage enzyme (P450scc) raised in the rabbit (1:200, AB1244; Chemicon International, Temecula, CA, USA), monoclonal anti-human cytochrome P450 aromatase (P450arom) raised in the mouse (1:100, MCA2077; Serotec, Oxford, UK) or polyclonal anti-human tartrate-resistant acid phosphatase (TRAP; also called type 5 acid phosphatase, ACP5) raised in the rabbit in order to detect uteroferrin (1:100, CN6911; ProSci, Poway, CA, USA) at 4 C in a moisture chamber.

Techniques: Immunostaining

Journal: Oncotarget

Article Title: Mitochondrial targeted HSP90 inhibitor Gamitrinib-TPP (G-TPP) induces PINK1/Parkin-dependent mitophagy

doi: 10.18632/oncotarget.22287

Figure Lengend Snippet: ( A , B ) G-TPP leads to Ub-charging of the active site mutant Parkin C431S. HeLa cells stably expressing 3xFLAG-Parkin C431S were treated with 10 µM G-TPP for the indicated times. As positive control some cells were treated with 10 µM CCCP for 2 h. (A) Western blots were prepared with cell lysates and probed with antibodies against FLAG. Ub-charged Parkin can be observed as 8 kDa band shift that collapses again with NaOH treatment. (B) Densitometric quantification of Parkin Ub-charging normalized to 2 h CCCP treatment. Shown is the average of three independent experiments ± SEM (one-way ANOVA with Tukey’s post-hoc test, *** p < 0.0005). ( C ) HeLa cells stably expressing untagged Parkin were treated with G-TPP for the indicated times. Western blots were prepared with cell lysates and probed with antibodies against Parkin substrates (Mitofusin 1 and 2, TOM70 and Miro1). Levels of Parkin substrates initially decreased but then recovered during the time course. Levels of TRAP1 were unchanged upon G-TPP treatment.

Article Snippet: The following antibodies have been used for western blot (WB) or immunofluorescence (IF): beta III tubulin (#5568, CST, WB: 1/2,000 or AB9354, Millipore, IF: 1/250), FLAG (F3165, Sigma, WB: 1/150,000), GAPDH (H86504M, Meridian Life Sciences, WB: 1/500,000), LC3B (NB100-2220, Novus Biologicals, WB: 1/5,000), Miro1 (H00055288-M01, Novus Biologicals, WB: 1/500), Mitofusin 1 (ab57602, Abcam, WB: 1/5,000), Mitofusin 2 (ab56889, Abcam, WB: 1/5,000), NBR1 (H00004077-M01, Abnova, WB: 1/500, IF: 1/100), NDP52 (12229-1-AP, PTG, WB: 1/1,000, IF: 1/400), OPTN (sc-166576, Santa Cruz, IF: 1/100), OPTN (10837-1-AP, PTG, WB: 1/5,000), p38 (#9212, CST, WB: 1/2,000), p62 (610832, BD Biosciences, WB: 1/2,000, IF:1/500), Parkin (#4211, CST, WB: 1/3,000), PGAM5 (ab12653, Abcam, WB: 1/5,000), PINK1 (#6946, CST, WB: 1/2,000, IF: 1/1,000), PINK1 (BC100-494, Novus Biologicals, WB: 1/2,000), TAX1BP1 (#5105, CST, WB: 1/2,000, IF: 1/400), TBK1 (#3504, CST, WB: 1/1,000), pS172-TBK1 (#5483, CST, WB: 1/1,000), TOM20 rabbit (11802-1-AP, PTG, IF: 1/2,000), TOM20 mouse (sc-17764, Santa Cruz, IF: 1/100), TOM70 (14528-1-AP, PTG, WB: 1/5,000), TRAP1 (#13405, CST, WB: 1/1,000), ubiquitin (#3933, CST, WB: 1/2,000), pS65-Ub (in-house [ , ], WB: 1/15,000, IF: 1/250), VDAC1 (ab14734, Abcam, WB: 1/10,000), vinculin (V9131, Sigma, WB: 1/500,000).

Techniques: Mutagenesis, Stable Transfection, Expressing, Positive Control, Western Blot, Electrophoretic Mobility Shift Assay

Journal: Poultry Science

Article Title: MiR-223 inhibits proliferation and steroid hormone synthesis of ovarian granulosa cell via the AKT signaling pathway by targeting CRIM1 in chicken

doi: 10.1016/j.psj.2024.103910

Figure Lengend Snippet: Antibody dilution concentration.

Article Snippet: anti-CYP19A1 , ABclonal (Wuhan, China) , A12684 , 1:800.

Techniques: Concentration Assay

Journal: ASN NEURO

Article Title: Sex-Dimorphic Octadecaneuropeptide (ODN) Regulation of Ventromedial Hypothalamic Nucleus Glucoregulatory Neuron Function and Counterregulatory Hormone Secretion

doi: 10.1177/17590914231167230

Figure Lengend Snippet: Description of Primary Antisera Used.

Article Snippet: Aromatase , Novus Biologicals LLC, Centennial CO, US , NB100-1596 , AB_10000919 , WB; 1:2000 (0.5 uL/mL) , Human Aromatase protein (between residues 400-502). [UniProt# P11511] , Giles et al., Breast Cancer Res. 2018; 20(1): 50 , 29898754.

Techniques: Recombinant, Sequencing

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: A novel variant of ER-alpha, ER-alpha36 mediates testosterone-stimulated ERK and Akt activation in endometrial cancer Hec1A cells

doi: 10.1186/1477-7827-7-102

Figure Lengend Snippet: Letrozole inhibits ER-α36-mediated ERK and Akt phosphorylation . Hec1A cells were pre-treated with 10 nM aromatase inhibitor letrozole (Lanes 3 and 4) for 1 h and then the cells were treated with vehicle (DMSO) (Lanes 1) and 10 nM testosterone (Lanes 2 and 4) for 5 min. Phospho-ERK1/2 (A) or phospho-Akt (B) were examined with specific antibodies. The same blot was stripped and probed with anti-total ERK or Akt antibodies. Hec1A cells were treated with 10 nM testosterone (Lanes 2) or together with 10 nM aromatase inhibitor letrozole (Lanes 3) overnight, and then aromatase expression was detected by Western blot using specific antibody against aromatase (C).

Article Snippet: Anti-aromatase antibody was purchased from Novus Biologicals (Novus Biologicals, Littleton, CO).

Techniques: Phospho-proteomics, Expressing, Western Blot

Journal: Journal of Virology

Article Title: Testicular Degeneration and Infertility following Arbovirus Infection

doi: 10.1128/JVI.01131-18

Figure Lengend Snippet: 3β-HSD and P450 expression in BTV-1 experimentally infected rams and mock-infected controls. (A and B) Immunohistochemistry showing expression of 3β-HSD (A) and P450 (B) (in brown) by the Leydig cells in sections of the testes of mock-infected healthy control rams. Arrows point to Leydig cells. (C to F) Immunohistochemistry showing lack of expression of 3β-HSD (C and E) and P450 (D and F) by Leydig cells in the testes of rams infected with BTV-1 IT2006 (C and D) or BTV-1 IT2013 (E and F). The peritubular areas where the Leydig cells are located are highlighted with a broken line. Scale bars, 100 μm. (G) Graph representing the relative levels of testosterone, assessed by ELISA, in the blood of rams infected with either BTV-1 IT2006 or BTV-1 IT2013 . Values are shown as percentages of the values of testosterone taken at day 0 before virus infection. Note the significant reduction in the levels of testosterone in rams infected with BTV-1 IT2006 between 0 and 8 to 11 dpi ( P < 0.05; one-way ANOVA) and in rams infected with BTV-1 IT2013 between 0 and either 5 or 15 dpi ( P < 0.05; one-way ANOVA).

Article Snippet: Sections were incubated overnight at 4°C using primary antibodies specific against the following markers: BTV NS2 ( , ), vimentin (Dako Agilent), von Willebrand factor (Dako Agilent), CD3 (Dako Agilent), MX-1 ( , ), 3β-HSD (Santa Cruz Biotechnology), P450 Aromatase (AcrisAntibodies GmbH), inhibin α (Ventana Medical Systems), smooth muscle actin (Dako Agilent), and KI-67 (Dako Agilent).

Techniques: Expressing, Infection, Immunohistochemistry, Control, Enzyme-linked Immunosorbent Assay, Virus

Journal: PLoS ONE

Article Title: Effect of Testosterone on Neuronal Morphology and Neuritic Growth of Fetal Lamb Hypothalamus-Preoptic Area and Cerebral Cortex in Primary Culture

doi: 10.1371/journal.pone.0129521

Figure Lengend Snippet: Shown is immunofluorescence labeling for androgen receptor (AR) and aromatase (Arom). Scale Bar = 50 μm.

Article Snippet: Immunocytochemistry was performed using the following primary antibodies: a mouse monoclonal anti-neuronal class III β-tubulin antibody (diluted 1:1000, Covance TUJ1, Covance Inc.; Princeton, NJ), PG-21 rabbit polyclonal anti-androgen receptor antibodies (1:100, Upstate 06–680; EMD Millipore; Billerica, MA) and rabbit polyclonal anti-aromatase antibodies (1:20, Acris Antibodies Inc, SanDiego, CA).

Techniques: Immunofluorescence, Labeling

Journal: The Journal of steroid biochemistry and molecular biology

Article Title: Effect of sildenafil on human aromatase activity: From in vitro structural analysis to catalysis and inhibition in cells.

doi: 10.1016/j.jsbmb.2016.09.003

Figure Lengend Snippet: Figure 1. UV-vis spectroscopy shows a Type II binding of sildenafil to human aromatase. A) Visible direct

Article Snippet: Confluent adherent cells grown in 10 cm diameter dishes were transiently transfected in Opti-MEM reduced serum medium (Gibco) by mixing 10 μg of the expression vector pCMV6-XL4 carrying the cDNA coding for the full-length aromatase (Origene, Rockville, USA) or a control empty vector (pIRES-puro2) and 10 μL of Lipofectamine2000 (Invitrogen) according to the manufacturer recommendations.

Techniques: UV-Vis Spectroscopy, Binding Assay