Journal: Archives of Gynecology and Obstetrics

Article Title: Glycosylated fibronectin as a first trimester marker for gestational diabetes

doi: 10.1007/s00404-020-05670-8

Figure Lengend Snippet: a Glycosylated fibronectin concentrations in BMI subgroups of 20–25 and 30–35 kg/m 2 in the study and the control group. b Effect of smoking on concentrations of glycosylated fibronectin in the control and GDM groups. c Multiple of median (MoM) values for fibronectin and glycosylated fibronectin between control and GDM groups. d Multiple of median (MoM) values for adiponectin and glycosylated adiponectin between control and GDM groups

Article Snippet: Calibrators for both assays were made from recombinant human adiponectin (1065-AF, R&D Systems, Abingdon, UK).

Techniques: Control

Journal: Diabetes

Article Title: Adiponectin Reduces Plasma Triglyceride by Increasing VLDL Triglyceride Catabolism

doi: 10.2337/db07-0435

Figure Lengend Snippet: Elevated adiponectin has no effect on HNF4α protein levels in mouse liver. Three days after viral vector injection, liver tissues were collected in the fasting state after perfusion. Total liver protein was prepared, and HNF4α and adiponectin were analyzed by Western blotting.

Article Snippet: Recombinant human adiponectin and anti-mouse adiponectin antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Plasmid Preparation, Injection, Western Blot

Journal: Human reproduction (Oxford, England)

Article Title: Expression of adiponectin receptors and effects of adiponectin isoforms in mouse preimplantation embryos.

doi: 10.1093/humrep/deq193

Figure Lengend Snippet: Figure 1 Qualitative RT–PCR analysis of adiponectin receptors (AdipoR1, AdipoR2) and adiponectin in oocytes and preimplantation embryos. Agarose gels with separated PCR products are shown. Lanes: Oc, oocytes; 4c, 4-cell embryos; 8c, 8- to16-cell embryos; Mo, morulas; Bl, blastocysts; MW, molecular weight markers; P, posi- tive control tissue (brain for adiponectin receptors, adipose tissue for adiponectin). The sizes of the MW and the predicted sizes of the PCR products in base pairs (bp) are indicated to the right and the left of the panels, respectively.

Article Snippet: Embryo culture and morphological evaluation Four-cell stage embryos were flushed from the oviduct, then pooled, randomly divided into two groups, washed in KSOM culture medium (Specialty Media Group, Phillipsburg, NJ, USA), transferred to KSOM culture drops (1 embryo/1 ml KSOM, covered with mineral oil) containing adiponectin (experimental group) or equivalent amounts of solvent (control group), and cultured in standard conditions (5% CO2 and 378C) for 65 h. Three isoforms of adiponectin were used in the experiments: fulllength adiponectin (mouse Acrp30, recombinant protein, produced in HEK 293 cells, GenWay, San Diego, CA, USA), the truncated isoform—globular adiponectin (mouse Acrp30, globular recombinant protein, produced in Escherichia coli, GenWay), and mutated full-length adiponectin (mouse Acrp30, trimeric recombinant protein, produced in HEK 293 cells, GenWay) with cystein 39 replaced with alanine (enabling trimer formation only).

Techniques: Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Control

Journal: Human reproduction (Oxford, England)

Article Title: Expression of adiponectin receptors and effects of adiponectin isoforms in mouse preimplantation embryos.

doi: 10.1093/humrep/deq193

Figure Lengend Snippet: Figure 4 Mean cell number in blastocysts after incubation with three different adiponectin isoforms. The 4-cell mouse embryos were incubated in the presence of adiponectin (FL AD, full-length adi- ponectin; GL AD, globular adiponectin; TR AD, trimeric adiponectin) for 65 h, and then the cell number in embryos that reached the blas- tocyst stage was determined. Values are arithmetical means + SEM. Black columns, adiponectin-treated groups; white columns, corre- sponding control groups. Total number of evaluated blastocysts in the experimental groups: FL AD 133, CTRL 160; GL AD 148, CTRL 156; TR AD 141, CTRL 139. Statistical difference between adiponectin-treated groups and corresponding control groups was assessed with unpaired Student’s t-test: **P , 0.01.

Article Snippet: Embryo culture and morphological evaluation Four-cell stage embryos were flushed from the oviduct, then pooled, randomly divided into two groups, washed in KSOM culture medium (Specialty Media Group, Phillipsburg, NJ, USA), transferred to KSOM culture drops (1 embryo/1 ml KSOM, covered with mineral oil) containing adiponectin (experimental group) or equivalent amounts of solvent (control group), and cultured in standard conditions (5% CO2 and 378C) for 65 h. Three isoforms of adiponectin were used in the experiments: fulllength adiponectin (mouse Acrp30, recombinant protein, produced in HEK 293 cells, GenWay, San Diego, CA, USA), the truncated isoform—globular adiponectin (mouse Acrp30, globular recombinant protein, produced in Escherichia coli, GenWay), and mutated full-length adiponectin (mouse Acrp30, trimeric recombinant protein, produced in HEK 293 cells, GenWay) with cystein 39 replaced with alanine (enabling trimer formation only).

Techniques: Incubation, Control

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 1. Viability of BME-UV1 bovine mammary epithelial cells treated for 24 h with different concentrations of chemerin, leptin, or adiponectin. Graphs present cell viability measured using the MTT assay. The viability of untreated control cells (0) was designated as 100%. Results are presented as means ± standard deviation of three independent experiments.

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: MTT Assay, Control, Standard Deviation

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 2. Expression of adipokine receptors in BME-UV1 bovine mammary epithelial cells. (A,B) Expression of adiponectin receptors (ADIPOR1 and ADIPOR2); (C,D) expression of chemerin receptors (CMLKR1 and GPR1). The relative mRNA expression of the analyzed genes was normalized to the mean expression of the histone reference gene. The expression of each analyzed gene in cells cultured in control medium (Ctrl.) was given as 1. Treatment with adiponectin at concentrations of 10 ng/mL or 500 ng/mL is marked as A10 and A500, respectively. Treatment with chemerin at concentrations of 10 ng/mL or 100 ng/mL is marked as Ch10 and Ch100, respectively. Results are presented as means ± standard deviation of three independent experiments performed in duplicate. Values that differed significantly from control are marked as ** (p < 0.01) or *** (p < 0.001).

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Expressing, Cell Culture, Control, Standard Deviation

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 4. Evaluation of apoptosis in BME-UV1 bovine mammary epithelial cells treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL) for 24 h. (A) Representative dot-plots of Annexin V/PI double staining in untreated control cells; (B) graph showing percentage of apoptotic cells (sum of Annexin Vpos/PIneg and Annexin Vpos/Pipos cells) in control and adipokine-treated cells; (C) representative images of Western blot (WB) analysis of apoptotic markers cleaved caspase 3 and bax, where gapdh was used as a reference protein; (D,E) graphs showing the results of the densitometric analysis of WB images; the integrated optical density (IOD) of cleaved caspase 3 and bax bands was normalized to the IOD of gapdh bands. Results are presented as means ± standard deviation of three or four independent experiments.

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Double Staining, Control, Western Blot, Standard Deviation

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 5. Concentration of αS1-casein in BME-UV1 cells (A) and media (B) collected after 24 h of culture. Cell were treated with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL) or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Un- treated cells were used as a negative control, whereas cells exposed for 24 h to the lactogenic hormone prolactin (PRL, 1 µg/mL) were used as a positive control. Results are presented as means ± standard deviation of three independent experiments. Means followed by a common letter are not significantly different according to one-way ANOVA with Tukey’s multiple comparison post-test, at the 5% level of significance (p < 0.05).

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Concentration Assay, Negative Control, Positive Control, Standard Deviation, Comparison

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 6. Panels of confocal micrographs presenting the immunofluorescence staining of BME-UV1 cells cultured on Matrigel for 11 days in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). Cells were stained with primary antibodies against β1-integrin and secondary antibodies conjugated with Alexa Fluor 488 dye (green fluorescence); Alexa Fluor 594 phalloidin, detecting F-actin (red fluorescence), and nuclei counterstained with Hoechst 33342 (blue fluorescence). Images were taken at 600× magnification and are representative for three independent experiments. Scale bar: 20 µm.

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Immunofluorescence, Staining, Cell Culture, Control, Fluorescence

Journal: International journal of molecular sciences

Article Title: Chemerin Stimulates the Secretory Activity of BME-UV1 Bovine Mammary Epithelial Cells.

doi: 10.3390/ijms25084147

Figure Lengend Snippet: Figure 7. Diameters of 3D mammospheres formed by BME-UV1 cells cultured on Matrigel for 11 days. Cells were cultured in control growth medium (Ctrl.) or medium supplemented with chemerin (Ch10 = 10 ng/mL, Ch100 = 100 ng/mL), leptin (L10 = 10 ng/mL, L100 = 100 ng/mL), or adiponectin (A10 = 10 ng/mL, A500 = 500 ng/mL). The diameters of cross sections of the spheroids were measured using ImageJ software (https://ij.imjoy.io accessed on 2 April 2024). Results are presented as means ± standard deviation of at least 14 spheroids per experimental condition photographed using a confocal microscope equipped with a digital camera.

Article Snippet: Chemerin (cat.no: 2325-CM-025), leptin (cat.no: 498-OB-01M), adiponectin (cat.no: 5095-AC-050), and prolactin (cat. no: 682-PL-050) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Control, Software, Standard Deviation, Microscopy

Journal: Molecular Psychiatry

Article Title: Adiponectin regulates contextual fear extinction and intrinsic excitability of dentate gyrus granule neurons through AdipoR2 receptors

doi: 10.1038/mp.2016.58

Figure Lengend Snippet: Adiponectin regulates contextual fear extinction. ( a ) Mice were subjected to fear conditioning (FC) followed by extinction training for 4 days (E1–E4). Adipo −/− mice showed normal contextual fear acquisition (left; genotype: F (1,24) =1.842, P =0.187; shock: F (4,96) =208.6, P <0.001; genotype × shock interaction: F (4,96) =1.354, P =0.256) and retrieval (middle; t (24) =0.225, P =0.824) but displayed slower extinction (right, genotype: F (1,24) =8.017, P =0.009; day: F (3,72) =101.6, P <0.001; genotype × day interaction: F (3,72) =2.739, P =0.0496). Adipo −/− , n =14; wild type (WT), n =12. *P< 0.05 and ** P <0.01 compared with WT littermate controls. ( b ) After contextual FC, dentate gyrus (DG)-cannulated WT mice received intra-DG infusions of adiponectin or vehicle (artificial cerebrospinal fluid (aCSF)) for 3 days, followed by extinction training for 4 days (E1–E4). Intra-DG infusion was given daily 30 min before re-exposure to the context during extinction training. In fear-conditioned mice (left), intra-DG infusions of adiponectin did not affect fear retrieval ( t (19) =0.238, P =0.815; middle left) but facilitated fear extinction (middle right; treatment: F (1,19) =3.422, P =0.08; day: F (3,57) =17.6, P <0.001; treatment × day interaction F (3,57) =2.892, P =0.043). Right, microinjection sites in the DG (open circles for vehicle-treated mice and closed circles for adiponectin-treated mice). Intra-DG vehicle (Veh), n =11; intra-DG adiponectin (Adipo), n =10. * *P< 0.01, # P =0.057 compared with the vehicle-treated group. Error bars represent s.e.m.

Article Snippet: Recombinant mouse adiponectin (R&D Systems, Minneapolis, MN, USA) was dissolved in artificial cerebrospinal fluid (137 m M NaCl, 2.7 m M KCl, 0.5 m M MgCl 2 ·6H 2 O, 0.9 m M CaCl 2 ·2H 2 O, 1.5 m M KH 2 PO 4 and 8.1 m M Na 2 HPO 4 ).

Techniques: Microinjection

Journal: Molecular Psychiatry

Article Title: Adiponectin regulates contextual fear extinction and intrinsic excitability of dentate gyrus granule neurons through AdipoR2 receptors

doi: 10.1038/mp.2016.58

Figure Lengend Snippet: Adiponectin modulates intrinsic excitability of dentate gyrus (DG) granule neurons. ( a ) Increased excitability of DG neurons in Adipo −/− mice. ( a1 ) Left, action potential (AP) trains induced by injecting a 90-pA current. The shaded area indicating the first 300 ms. Right, number of APs (genotype: F (1,19) =5.484, P =0.03; current: F (6,114) =77.18, P <0.001; genotype × current interaction: F (6,114) =3.113, P =0.007); ( a2 ) rheobase current ( t (19) =1.873, P =0.076); ( a3 ) resting membrane potential ( t (19) =0.175, P =0.863); ( a4 ) input resistance ( t (19) =2.632, P =0.0164); ( a5 ) AP threshold ( t (19) =0.475, P =0.640). n =10 from 3 wild-type (WT) mice; n =11 from 3 Adipo −/− mice. * P< 0.05, # P =0.076, **P <0.01 and ***P <0.001 compared with WT littermates. ( b ) Decreased excitability of DG neurons in WT mice after bath application of 15 μ M AdipoRon. ( b1 ) Left, action potential trains elicited by injecting a 90-pA current. Right, number of AP (treatment: F (1,9) =8.757, P =0.016; current: F (6,54) =34.61, P <0.001; treatment × current interaction: F (6,54) =7.338, P <0.001); ( b2 ) rheobase current ( t (9) =2.449, P =0.0368); ( b3 ) resting membrane potential ( t (9) =3.496, P =0.0068); ( b4 ) input resistance ( t (9) =1.533, P =0.159); ( b5 ) AP threshold ( t (9) =0.235, P =0.819). n =10 from 4 mice. * P< 0.05, ** P< 0.01 and *** P< 0.001 compared with artificial cerebrospinal fluid (aCSF) treatment. ( c ) Decreased excitability of DG neurons in Adipo −/− mice by bath application of 15 μ M AdipoRon. ( c1 ) Left, action potential trains induced by injecting a 90-pA current. Right, number of APs (treatment: F (1,8) =8.475, P =0.02; current: F (6,48) =24.30, P <0.001; treatment × current interaction: F (6,48) =5.846, P <0.001); ( c2 ) rheobase current ( t (8) =3.500, P =0.0081); ( c3 ) resting membrane potential ( t (8) =5.487, P =0.0006); ( c4 ) input resistance ( t (8) =1.688, P =0.130); ( c5 ) AP threshold ( t (8) =1.294, P =0.232). n =9 from 4 mice. ** P< 0.01 and *** P< 0.001 compared with aCSF treatment. Error bars represent s.e.m.

Article Snippet: Recombinant mouse adiponectin (R&D Systems, Minneapolis, MN, USA) was dissolved in artificial cerebrospinal fluid (137 m M NaCl, 2.7 m M KCl, 0.5 m M MgCl 2 ·6H 2 O, 0.9 m M CaCl 2 ·2H 2 O, 1.5 m M KH 2 PO 4 and 8.1 m M Na 2 HPO 4 ).

Techniques: Membrane

Journal: Molecular Psychiatry

Article Title: Adiponectin regulates contextual fear extinction and intrinsic excitability of dentate gyrus granule neurons through AdipoR2 receptors

doi: 10.1038/mp.2016.58

Figure Lengend Snippet: AdipoR2 knockout causes abnormal contextual fear responses and resistance to the facilitating effect of intra-dentate gyrus (DG) adiponectin on contextual fear extinction. ( a ) AdipoR2 −/− mice showed normal contextual fear acquisition (genotype: F (1,20) =1.292, P =0.269; shock: F (4,80) =159.1, P <0.001; genotype × shock interaction: F (4,80) =0.679, P =0.609), but elevated levels of freezing during fear retrieval ( t (20) =3.648 P =0.0016) and fear extinction (Freezing time; genotype: F (1,20) =38.16, P <0.001; day: F (3,60) =130.0, P <0.001; genotype × day interaction: F (3,60) =2.043, P =0.118) and slower extinction rate (% of initial freezing; genotype: F (1,20) =5.895, P =0.0247; day: F (3,60) =126.9, P <0.001; genotype × day interaction: F (3,60) =3.102, P =0.033). Wild type (WT), n =11; AdipoR2 −/− , n =11. * P< 0.05, ** P< 0.01 and *** P< 0.001 compared with WT. ( b ) AdipoR2 −/− mice and WT mice bilaterally cannulated in the DG showed similar levels of freezing during contextual fear conditioning (genotype: F (1,19) =1.626, P =0.218; shock: F (4,76) =95.7, P <0.001; genotype × shock interaction: F (4,76) =2.218, P =0.075). Subsequently, the WT mice received intra-DG infusions of vehicle (WT-Veh, n =7) and AdipoR2 −/− mice received intra-DG infusions of vehicle (AdipoR2 −/− -Veh, n =6) or adiponectin (AdipoR2 −/− -Adipo, n =8) for 3 days, followed by 4 days of extinction training (E1–E4) with daily intra-DG infusion 30 min before re-exposure to the context. AdipoR2 −/− mice displayed increased levels of freezing during contextual fear retrieval (F (2,18) =4.940, P =0.0195) and extinction (Freezing time; treatment: F (2,18) =12.083, P <0.001; day: F (3,54) =55.34, P <0.001; treatment × day interaction: F (6,54) =1.358, P =0.248). There was no difference in extinction rates between three treatment groups (% initial freezing; treatment: F (2,18) =3.344, P =0.058; day: F (3,54) =53.36, P <0.001; treatment × day interaction: F (6,54) =1.831, P =0.110). * P< 0.05 and ** P< 0.01 compared with the WT-Veh group. * P< 0.05 and ** P< 0.01, AdipoR2 −/− -Veh versus WT-Veh; # P< 0.05, ### P< 0.001, AdipoR2 −/− -Adipo versus WT-Veh. Error bars represent s.e.m.

Article Snippet: Recombinant mouse adiponectin (R&D Systems, Minneapolis, MN, USA) was dissolved in artificial cerebrospinal fluid (137 m M NaCl, 2.7 m M KCl, 0.5 m M MgCl 2 ·6H 2 O, 0.9 m M CaCl 2 ·2H 2 O, 1.5 m M KH 2 PO 4 and 8.1 m M Na 2 HPO 4 ).

Techniques: Knock-Out

Journal: Biology

Article Title: Estrogenic Compounds or Adiponectin Inhibit Cyclic AMP Response to Human Luteinizing Hormone in Mouse Leydig Tumor Cells

doi: 10.3390/biology8020045

Figure Lengend Snippet: Effect of adiponectin (ADPN) on the cAMP response to 0.7 nM hLH in Mouse Leydig Tumor cells (mLTC)-1 cells. ( A ): Kinetics of oxyluciferin luminescence produced by cyclic AMP-dependent luciferase in mLTC-1 cells stimulated by 0.7 nM rec hLH after 1h-preincubation with the shown concentrations of ADPN. ( B ): Dose-response effects from kinetics in panel (A). ( C ) Dose-response effect of 1h-preincubation with frozen and thawed ADPN on the cAMP response to 0.7 nM hLH. Each point represents the mean of luminescence in six wells in each condition. The figure presents one representative experiment out of three independent experiments. After checking of normality of distribution, the data were analyzed by a paired Student’s t-test. * indicates a significant difference (* p < 0.05, *** p < 0.001) compared between each ADPN concentration and control (hLH alone), ns = not statistically significant.

Article Snippet: Recombinant bovine adiponectin was purchased from Cusabio (Interchim, Montluçon, France).

Techniques: Produced, Luciferase, Concentration Assay, Control

Journal: Cardiology Plus

Article Title: Effect of Exogenous Adiponectin on Myocardial Fibrosis in Mice Models with Myocardial Fibrosis

doi: 10.4103/2470-7511.248369

Figure Lengend Snippet: Figure 1: Sirius red‑stained section of myocardium. Upper: sham (n = 10); middle: Isoproterenol + adiponectin (n = 10); bottom: isoproterenol (n = 10). Left column: optical microscopy; right column: polarized light microscopy (×20)

Article Snippet: From day 4, mice in the APN + ISO group underwent intraperitoneal injection of murine recombinant adiponectin (10 μg/kg • d; Cusabio Biotech, Wuhan, China), whereas mice in the sham and ISO groups were given normal saline by intraperitoneal injection for 32 days.

Techniques: Microscopy, Light Microscopy

Journal: Cardiology Plus

Article Title: Effect of Exogenous Adiponectin on Myocardial Fibrosis in Mice Models with Myocardial Fibrosis

doi: 10.4103/2470-7511.248369

Figure Lengend Snippet: Figure 2: Western blot analyses of type‑I and type‑III collagen protein in the myocardium. Upper: type‑I collagen protein; bottom: type‑III collagen protein. From the left, lane 1 and 2 show the electrophoretogram of the sham group, lanes 3–7 represent the electrophoretogram of the isoproterenol group, and lanes 8–12 show the electrophoretogram of the isoproterenol + adiponectin group

Article Snippet: From day 4, mice in the APN + ISO group underwent intraperitoneal injection of murine recombinant adiponectin (10 μg/kg • d; Cusabio Biotech, Wuhan, China), whereas mice in the sham and ISO groups were given normal saline by intraperitoneal injection for 32 days.

Techniques: Western Blot

Journal: Human reproduction (Oxford, England)

Article Title: Expression of adiponectin receptors and effects of adiponectin isoforms in mouse preimplantation embryos.

doi: 10.1093/humrep/deq193

Figure Lengend Snippet: Figure 1 Qualitative RT–PCR analysis of adiponectin receptors (AdipoR1, AdipoR2) and adiponectin in oocytes and preimplantation embryos. Agarose gels with separated PCR products are shown. Lanes: Oc, oocytes; 4c, 4-cell embryos; 8c, 8- to16-cell embryos; Mo, morulas; Bl, blastocysts; MW, molecular weight markers; P, posi- tive control tissue (brain for adiponectin receptors, adipose tissue for adiponectin). The sizes of the MW and the predicted sizes of the PCR products in base pairs (bp) are indicated to the right and the left of the panels, respectively.

Article Snippet: Embryo culture and morphological evaluation Four-cell stage embryos were flushed from the oviduct, then pooled, randomly divided into two groups, washed in KSOM culture medium (Specialty Media Group, Phillipsburg, NJ, USA), transferred to KSOM culture drops (1 embryo/1 ml KSOM, covered with mineral oil) containing adiponectin (experimental group) or equivalent amounts of solvent (control group), and cultured in standard conditions (5% CO2 and 378C) for 65 h. Three isoforms of adiponectin were used in the experiments: fulllength adiponectin (mouse Acrp30, recombinant protein, produced in HEK 293 cells, GenWay, San Diego, CA, USA), the truncated isoform—globular adiponectin (mouse Acrp30, globular recombinant protein, produced in Escherichia coli, GenWay), and mutated full-length adiponectin (mouse Acrp30, trimeric recombinant protein, produced in HEK 293 cells, GenWay) with cystein 39 replaced with alanine (enabling trimer formation only).

Techniques: Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Control

Journal: Human reproduction (Oxford, England)

Article Title: Expression of adiponectin receptors and effects of adiponectin isoforms in mouse preimplantation embryos.

doi: 10.1093/humrep/deq193

Figure Lengend Snippet: Figure 4 Mean cell number in blastocysts after incubation with three different adiponectin isoforms. The 4-cell mouse embryos were incubated in the presence of adiponectin (FL AD, full-length adi- ponectin; GL AD, globular adiponectin; TR AD, trimeric adiponectin) for 65 h, and then the cell number in embryos that reached the blas- tocyst stage was determined. Values are arithmetical means + SEM. Black columns, adiponectin-treated groups; white columns, corre- sponding control groups. Total number of evaluated blastocysts in the experimental groups: FL AD 133, CTRL 160; GL AD 148, CTRL 156; TR AD 141, CTRL 139. Statistical difference between adiponectin-treated groups and corresponding control groups was assessed with unpaired Student’s t-test: **P , 0.01.

Article Snippet: Embryo culture and morphological evaluation Four-cell stage embryos were flushed from the oviduct, then pooled, randomly divided into two groups, washed in KSOM culture medium (Specialty Media Group, Phillipsburg, NJ, USA), transferred to KSOM culture drops (1 embryo/1 ml KSOM, covered with mineral oil) containing adiponectin (experimental group) or equivalent amounts of solvent (control group), and cultured in standard conditions (5% CO2 and 378C) for 65 h. Three isoforms of adiponectin were used in the experiments: fulllength adiponectin (mouse Acrp30, recombinant protein, produced in HEK 293 cells, GenWay, San Diego, CA, USA), the truncated isoform—globular adiponectin (mouse Acrp30, globular recombinant protein, produced in Escherichia coli, GenWay), and mutated full-length adiponectin (mouse Acrp30, trimeric recombinant protein, produced in HEK 293 cells, GenWay) with cystein 39 replaced with alanine (enabling trimer formation only).

Techniques: Incubation, Control