Journal: Scientific Reports

Article Title: Human Sertoli cells support high levels of Zika virus replication and persistence

doi: 10.1038/s41598-018-23899-x

Figure Lengend Snippet: Sertoli cells support high levels of ZIKV replication. ( A ) Primary human Sertoli and Leydig cells and the human A549 cell line were infected with ZIKV strain MR766 (MR) or PRVABC59 (PR) (MOI = 5) for 24, 48, 72 and 96 hours. At each time point, supernatants were harvested and viral titers were determined by plaque assay. ( B – E ) Cells were harvested at each time point and viral replication ( B ), Interferon-β ( C ), Interferon-λ2 ( D ) and OAS2 ( E ) mRNA levels were determined by qRT-PCR. ( F ) Sertoli cells seeded on coverslips were infected with ZIKV MR766 (MR) or PRVABC59 (PR) (MOI = 5) for 48 hours and then processed for indirect immunofluorescence using antibodies to ZIKV NS1 and GATA4 to identify infected cells expressing Sertoli markers. Nuclei were stained with DAPI. Representative images are shown. All values are expressed as mean ± standard error. N = 3.

Article Snippet: Primary human Leydig cells were purchased from ScienCell Research Laboratories, USA. and grown in Leydig Cell Medium (ScienCell Research Laboratories, USA).

Techniques: Infection, Plaque Assay, Quantitative RT-PCR, Immunofluorescence, Expressing, Staining

Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Role of the rate of internalization of the agonist-receptor complex on the agonist-induced down-regulation of the lutropin/choriogonadotropin receptor.

doi: 10.1210/mend.13.8.0331

Figure Lengend Snippet: Fig. 1. Agonist-Induced Internalization and Down-Regula- tion of the rLHR in Leydig Tumor Cells and in 293 Cells Stably Transfected with Different Truncations of the rLHR MA-10 and R2C cells, as well as clonal lines of 293 cells expressing the indicated truncations of the rLHR, were used. Down-regulation was measured after incubation of the cells with or without 100 ng/ml hCG for 24 h at 37 C. The cells were then washed to remove the free hormone, briefly treated with a pH 3 buffer to remove any receptor-bound hormone, and then used to measure [125I]hCG during a 4-h incubation at 4 C with 100 ng/ml [125I]hCG as described in Materials and Methods. The results of these experiments are shown in the top panel and are expressed as percent of the binding activity detected in cells incubated without hCG but otherwise treated under identical conditions. The t1/2 values of internal- ization of [125I]hCG were measured as described in Materials and Methods and are shown in the bottom panel. Each num- ber represents the average 6 SEM of three to five independent experiments. The asterisks indicate differences that are sta- tistically significant (P , 0.05) from 293L(wt-12) cells.

Article Snippet: Human kidney 293 cells and the rat Leydig tumor cell line (R2C) were purchased from the American Type Culture Collection.

Techniques: Stable Transfection, Transfection, Expressing, Incubation, Binding Assay, Activity Assay