Journal: Frontiers in Endocrinology

Article Title: Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

doi: 10.3389/fendo.2022.1010796

Figure Lengend Snippet: Gene expression of the mitogens Glial cell-line derived neurotrophic factor (GDNF) , Fibroblast growth factor 2 (FGF2) , and Stem cell factor (SCF) in Sertoli cells (SCs). RT-PCR analysis of (A) GDNF , (B) FGF2 , and (C) SCF . Data represent the mean ± SD of three independent experiments, each performed in triplicate. SCs were used as a positive control. Non-significant (NS) vs SCs, *p < 0.05 and **p < 0.001 vs SCs, ##p < 0.001 vs 0.33 ng/ml of rhIGF2, §§p < 0.001 vs 3.33 ng/ml, &p < 0.05 and &&p < 0.001 vs 10 ng/ml of rhIGF2, ns vs 10 ng/ml of rhIGF2.

Article Snippet: When SC monolayers were confluent (after 3 days of culture), they underwent the following treatments: (1) 0.33 ng/ml of rhIGF2 (Cayman Chemical, Ann Arbor, MI, USA) for 48 hours; (2) 3.33 ng/ml of rhIGF2 for 48 hours; (3) 10 ng/ml of rhIGF2 for 48 hours; (4) 1 μM of NVP-AE541 for 1 hour and then incubated with 0.33 ng/ml of rhIGF2 for 48 hours; (5) 1 μM of NVP-AE541 for 1 hour and then incubated with 3.33 ng/ml of rhIGF2 for 48 hours; (6) 1 μM of NVP-AE541 for 1 hour and then incubated with 10 ng/ml of rhIGF2 for 48 hours.

Techniques: Gene Expression, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Positive Control

Journal: Frontiers in Endocrinology

Article Title: Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

doi: 10.3389/fendo.2022.1010796

Figure Lengend Snippet: Anti-Müllerian hormone (AMH) and inhibin B gene expression and protein secretion. AMH mRNA and protein levels are shown in panels (A, B) , respectively. Inhibin B mRNA and protein levels are shown in panels (C, D) , respectively. Data represent the mean ± SD of three independent experiments, each performed in triplicate. Non-significant (NS) vs SCs, *p < 0.05 and **p < 0.001 vs SCs, ##p < 0.001 vs 0.33 ng/ml of rhIGF2, §§p < 0.001 vs 3.33 ng/ml, and &&p < 0.001 vs 10 ng/ml of rhIGF2.

Article Snippet: When SC monolayers were confluent (after 3 days of culture), they underwent the following treatments: (1) 0.33 ng/ml of rhIGF2 (Cayman Chemical, Ann Arbor, MI, USA) for 48 hours; (2) 3.33 ng/ml of rhIGF2 for 48 hours; (3) 10 ng/ml of rhIGF2 for 48 hours; (4) 1 μM of NVP-AE541 for 1 hour and then incubated with 0.33 ng/ml of rhIGF2 for 48 hours; (5) 1 μM of NVP-AE541 for 1 hour and then incubated with 3.33 ng/ml of rhIGF2 for 48 hours; (6) 1 μM of NVP-AE541 for 1 hour and then incubated with 10 ng/ml of rhIGF2 for 48 hours.

Techniques: Gene Expression

Journal: Frontiers in Endocrinology

Article Title: Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

doi: 10.3389/fendo.2022.1010796

Figure Lengend Snippet: FSHR gene and protein expression in Sertoli cells (SCs). (A) RT-PCR analysis of the FSHR gene. Data represent the mean ± SD of three independent Q3 experiments, each performed in triplicate. (B) Cytofluorimetric analysis. The bars represent the percentage of FSHR-positive cells without stimulation, after incubation with NVP-AEW541, IGF2, or NVP-AEW541 + IGF2. Compared with the control, the percentage of FSHR-positive cells was significantly reduced after incubation with IGF2 (2.82 ± 0.1% in the untreated controls, 1.87 ± 0.1% at 0.33 ng/mL, 2.19 ± 0.2% at 3.33 ng/mL, 2.36 ± 0.1% at 10 ng/mL; p < 0.01). The suppression was reversed after incubation with NVP- AEW541. Data represent the mean ± SD of three independent experiments, each performed in triplicate. FSHR, follicle-stimulating hormone receptor; IGF2, Insulin-like growth factor 2. *p < 0.05 and **p < 0.001 vs SCs, ##p < 0.001 vs 0.33 ng/ml of rhIGF2, §§p < 0.001 vs 3.33 ng/ml, &&p < 0.001 vs 10 ng/ml of rhIGF2”

Article Snippet: When SC monolayers were confluent (after 3 days of culture), they underwent the following treatments: (1) 0.33 ng/ml of rhIGF2 (Cayman Chemical, Ann Arbor, MI, USA) for 48 hours; (2) 3.33 ng/ml of rhIGF2 for 48 hours; (3) 10 ng/ml of rhIGF2 for 48 hours; (4) 1 μM of NVP-AE541 for 1 hour and then incubated with 0.33 ng/ml of rhIGF2 for 48 hours; (5) 1 μM of NVP-AE541 for 1 hour and then incubated with 3.33 ng/ml of rhIGF2 for 48 hours; (6) 1 μM of NVP-AE541 for 1 hour and then incubated with 10 ng/ml of rhIGF2 for 48 hours.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Incubation, Control

Journal: Frontiers in Endocrinology

Article Title: Sperm-carried IGF2 downregulated the expression of mitogens produced by Sertoli cells: A paracrine mechanism for regulating spermatogenesis?

doi: 10.3389/fendo.2022.1010796

Figure Lengend Snippet: Flow cytometric analysis of Sertoli cells stained with CFSE without stimulation, after incubation with IGF2, NVP-AEW541, and IGF2 + NVP-AEW541. *p < 0.05 vs SCs, #p < 0.05 vs 0.33 ng/ml of rhIGF2.

Article Snippet: When SC monolayers were confluent (after 3 days of culture), they underwent the following treatments: (1) 0.33 ng/ml of rhIGF2 (Cayman Chemical, Ann Arbor, MI, USA) for 48 hours; (2) 3.33 ng/ml of rhIGF2 for 48 hours; (3) 10 ng/ml of rhIGF2 for 48 hours; (4) 1 μM of NVP-AE541 for 1 hour and then incubated with 0.33 ng/ml of rhIGF2 for 48 hours; (5) 1 μM of NVP-AE541 for 1 hour and then incubated with 3.33 ng/ml of rhIGF2 for 48 hours; (6) 1 μM of NVP-AE541 for 1 hour and then incubated with 10 ng/ml of rhIGF2 for 48 hours.

Techniques: Staining, Incubation