Journal: Stem Cell Research & Therapy

Article Title: Insulin-like growth factor-II overexpression accelerates parthenogenetic stem cell differentiation into cardiomyocytes and improves cardiac function after acute myocardial infarction in mice

doi: 10.1186/s13287-020-1575-4

Figure Lengend Snippet: IGF-II overexpression promotes cardiomyocyte maturation in vivo. ESC- and PSC-derived cardiomyocytes at 1 × 10 5 cells/recipient mouse were injected into the heart at 1 week after acute myocardial infarction (MI). The IGF-II and VEGF levels in the injection sites ( a ) and the serum ( b ) were measured by ELISA at 4 weeks post-transplant. Data are expressed as the mean ± SD. * p < 0.05 vs. any other group. n = 12. c Immunofluorescence staining was performed to observe the cardiomyocyte morphology and to detect α-actinin expression (red staining) in the injection sites at 4 weeks post-transplant. Blue staining, Hoechst 33342. Green staining, eGFP. Scale bars, 10 μm

Article Snippet: IGF-II levels in serum (Quantikine ELISA Kit, cat# MG200, R&D Systems, Minneapolis, MN, USA), heart tissue (DuoSet ELISA, cat# DY792, R&D Systems), vascular endothelial growth factor (VEGF) levels in serum (cat# ab100751; Abcam), and heart tissue (cat# ab209882; Abcam) were determined with ELISA kits following the manufacturer’s instructions.

Techniques: Over Expression, In Vivo, Derivative Assay, Injection, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Expressing

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 1 Oocytes from obese mice display reduced serum IGF2 levels and reduced IGF2 mRNA expressions (A) Serum IGF2 contents in mice with normal diet (ND) and high-fat diet (HFD), measured by ELISA. n = 9 female mice for each group. B qPCR data indicating the mRNA expressions of IGF2 and target genes in mouse oocytes at different stages from ND and HFD mice. *p < 0.05. Student’s t-test (two-tailed) was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 2 Role of IGF2 in oocytes maturation of obese mice (A) Schematic diagram indicating treatment plan of oocytes in culture medium in vitro. B, C Percentage of GVBD and MII-stage oocytes from ND (n = 79) or HFD (n = 150) mice, and of oocytes from HFD mice given 50 nM IGF2 to the culture medium (“HFD + IGF2”). D Morphology of oocytes cultured in vitro and examined at the MII developmental stage. The arrows point to oocytes that are unable to transform into MII-stage. Scale bar, 100 μm. NS, not significant. One-way ANOVA test was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: In Vitro, Cell Culture

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 3 IGF2 administration improves the embryonic developmental competency of oocytes from obese mice (A) Schematic diagram indicating treatment plan of embryos in culture medium in vitro. B Quantitative analysis of morula and blastocyst in ND (n = 72), HFD (n = 57), and HFD+ IGF2 (n = 49). C Morphology of embryos cultured in vitro and examined at the blastocyst developmental stage. The arrows point to embryos that developed into blastocysts; the arrowheads showing the embryos which failed to transform into blastocysts. Scale bar, 300 μm. D Quantification of the total and TE number of cells in ND (n = 37), HFD (n = 20) and HFD + IGF2 (n = 26) blastocysts. E Representative images of blastocysts of the ND, HFD, and HFD + IGF2 groups. The trophectoderm (TE) was stained with CDX2 (green), and chromosomes were counterstained with DAPI (blue). Scale bar, 20 μm. NS, not significant. *p < 0.05. One-way ANOVA test was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: In Vitro, Cell Culture, Staining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 4 IGF2 ameliorates the quality of obese mice oocytes by reducing oxidative stress. A Morphology of spindle and chromosome organization in ND, HFD, and HFD + IGF2 oocytes. α-tubulin antibody (green) was used to stained the spindles while chromosomes were counterstained with Hoechst 33342 (blue). Scale bar = 10 μm. B Quantification of oocytes with spindle/chromosomes defects in ND (n = 38), HFD (n = 76), and HFD + IGF2 (n = 72) oocytes. C Quantification of ROS (DCFH-DA staining) signals in ND (n = 10), HFD, (n = 10), and HFD + IGF2 (n = 10) oocytes. D Representative confocal images showing CM-H2DCFDA fluorescence (green) in ND, HFD, and HFD + IGF2 oocytes. Scale bar = 20 μm. E Adenosine triphosphate (ATP) content in ND (n = 50), HFD (n = 50), and HFD + IGF2 (n = 50) oocytes. *p < 0.05. One-way ANOVA test was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: Staining, Fluorescence

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 5 IGF2 increases mitochondrial activity in oocytes from obese mice (A) ND, HFD, and HFD + IGF2 oocytes were stained with JC-1 to test the mitochondrial membrane potential (MMP). Scale bar = 10 μm. B Quantification of the ratio of red to green fluorescence intensity in ND (n = 20), HFD (n = 22), and HFD + IGF2 (n = 20) oocytes. C Mitochondria were labeled with mitotracker Green FM (green). Scale bar = 20 μm. D Quantification of mitochondrial distribution signals in ND (n = 148), HFD (n = 54), and HFD + IGF2 (n = 56) oocytes. E Immunofluorescence of HPG (green) showing the overall translation levels in MII-stage ND, HFD, and HFD + IGF2 oocytes. Chromosomes were counterstained with DAPI (blue). Scale bar = 20 μm. F HPG signal intensity quantification in ND (n = 16), HFD, (n = 20) and HFD + IGF2 (n = 17) oocytes. *p < 0.05. One-way ANOVA test was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: Activity Assay, Staining, Membrane, Fluorescence, Labeling, Immunofluorescence

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 6 IGF2 improves the mitochondrial ultrastructure of oocytes from obese mice (A) Mitochondrial micrograph of ND, HFD, and HFD + IGF2 oocytes at 5000x magnification examined by TEM. Scale bar = 1 μm. Note the normal mitochondria (Mn) and the vacuolated mitochondria (Mv). B Numbers of mitochondria per defined region of interest (ROI) in ND, HFD, and HFD + IGF2 oocytes. *p < 0.05. One-way ANOVA test was used for statistical analysis. Error bars indicate the SEM. C Mitochondrial micrograph from control and IGF2-treated oocytes at 60,000x magnification. Inner membrane (IM), outer membrane (OM), and intermembrane space (IMS). Scale bar = 100 nm

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: Control, Membrane

Journal: Reproductive biology and endocrinology : RB&E

Article Title: IGF2 reduces meiotic defects in oocytes from obese mice and improves embryonic developmental competency.

doi: 10.1186/s12958-022-00972-9

Figure Lengend Snippet: Fig. 7 IGF2 reduces apoptosis and induces expression of known target genes in oocytes from obese mice (A) Representative images of early apoptosis labeled with Annexin V of ND, HFD, and HFD + IGF2 oocytes. Red fluorescent indicates the Annexin V positive oocytes. Scale bar = 10 μm. B The percentage of early apoptosis in MII-stage oocytes from ND (n = 100),HFD (n = 64) and HFD+ IGF2 (n = 62) group. C qRT-PCR results showing IGF2 and target genes expression in mouse MII-stage oocytes from ND, HFD, and HFD + IGF2 groups after in vitro maturation. *p < 0.05. One-way ANOVA test and students t-test (two-tailed) was used for statistical analysis. Error bars indicate the SEM

Article Snippet: Serum IGF2 concentration was measured by following the protocol using an ELISA kit (RnD system, MG200).

Techniques: Expressing, Labeling, Quantitative RT-PCR, In Vitro, Two Tailed Test

Journal: bioRxiv

Article Title: Activation of proneuronal transcription factor Ascl1 in maternal liver ensures a healthy pregnancy

doi: 10.1101/2021.04.27.441617

Figure Lengend Snippet: Placentas were collected from gestation days (GD) 15 and 18 Ascl1 fl/fl and hep-Ascl1 -/- mice. (A) Frozen placental sections underwent periodic acid-Schiff (PAS) staining. Glycogen is stained red to purple. (B) Frozen placental sections were subjected to PL-I and PL-II in situ hybridization staining using RNAscope 2.5 HD Assay-BROWN kit. The PL-I and PL-II mRNAs are stained dark brown. (C) Quantification of relative intensity of Western blotting signals from . Data are presented as the mean fold changes relative to GD15 Ascl1 fl/fl mice (± SD; n = 3 for each group). *, P < 0.05, between Ascl1 fl/fl and hep-Ascl1 -/- mice. AKT, total protein kinase B; Ascl1 , achaete-scute homolog 1; ERK1/2, extracellular signal-regulated kinase 1/2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IGF2, insulin-like growth factor; PL-II, placental lactogen II.

Article Snippet: Serum IGF2 levels were quantified using a 1/6 dilution with the Mouse IGF-2 ELISA Kit (Boster Biological Technology, EK0381) as per directions by the manufacturer and read with the SpectraMax M2e spectrophotometer using the SoftMax Pro 6 program.

Techniques: Staining, In Situ Hybridization, RNAscope, HD Assay, Western Blot

Journal: bioRxiv

Article Title: Activation of proneuronal transcription factor Ascl1 in maternal liver ensures a healthy pregnancy

doi: 10.1101/2021.04.27.441617

Figure Lengend Snippet: Maternal livers were collected and weighed from nonpregnant (NP) and gestation days (GD) 15 and 18 Ascl1 fl/fl and hep-Ascl1 -/- mice. (A) Hepatic Igf2 mRNA levels were measured using qRT-PCR and presented as the mean fold changes relative to NP controls ± SD (n = 4-5). (B) Western blotting was performed using liver lysates with an antibody against IGF2. (C) Igf2 in situ hybridization on liver sections. (D) IGF2 immunostaining. (E) Levels of hepatic Igf2 promoter-specific transcript variants were measured using qRT-PCR and presented as the mean fold changes relative to NP controls ± SD (n = 4-5). (F) Levels of IGF2 protein in serum were measured using ELISA and presented as the mean fold changes ± SD (n = 3-5). *, P < 0.05; **, P < 0.01; ***, P < 0.001. P0, placental-specific Igf2 promoter; P1-3, placental- and fetal liver-specific Igf2 promoter.

Article Snippet: Serum IGF2 levels were quantified using a 1/6 dilution with the Mouse IGF-2 ELISA Kit (Boster Biological Technology, EK0381) as per directions by the manufacturer and read with the SpectraMax M2e spectrophotometer using the SoftMax Pro 6 program.

Techniques: Quantitative RT-PCR, Western Blot, In Situ Hybridization, Immunostaining, Enzyme-linked Immunosorbent Assay