r2c rat leydig tumor cells  (ATCC)

Journal: Reproductive toxicology (Elmsford, N.Y.)

Article Title: 3-chloro-1,2-propanediol induces oxidative stress and promotes testicular damage and infertility in rats through CYP2C9.

doi: 10.1016/j.reprotox.2024.108633

Figure Lengend Snippet: Fig. 6. 3-MCPD affects oxidative stress and promotes testicular cell apoptosis through CYP2C9. (A) The 2D structure of 3-MCPD. (B) Visualization result of molecular docking. (C) Western blot analysis of the CYP2C9 in R2C cells under control, low dose 3-MCPD (0.5 mmol/L), or high dose (2 mmol/L), (D) Western blot analysis of the CYP2C9 in R2C cells under control, high dose 3-MCPD (2 mmol/L) or 3-MCPD + sulphametoxazole (CYP2C9 inhibitor, SPZ) treatments for 24 hours. (E) Proliferation detection by CCK8 assay. (F) Progesterone synthesized quantitatively measured. (G) ROS levels using ELISA assay against R2C cell line. (H) Oxidative stress-related molecules HO-1 and SOD were detected by Western blot. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: R2C rat Leydig tumor cells (catalog no. CCL-97) was obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Western Blot, Control, CCK-8 Assay, Synthesized, Enzyme-linked Immunosorbent Assay

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 1 Effect of HEM on LC proliferation. MA-10 (A) and R2C (B) LC were incubated with increasing concentrations (1–25 µmol/L) of HEM or growth medium (GM) for 24 h. (C) Effect of HEM on normal ILCs proliferation. Rat ILCs were incubated with increasing concentrations of HEM (0.1– 25 µmol/L) or 0.5 mg/L IGF-1 for 24 h. The cells were labeled with [3H]-thymidine during the last 16 h of the incubation. Scale bars are means ± s.e.m. for a representative (n = 3) octuplicate experiment. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001 vs control.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, Labeling, Control

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 2 Effect of HEM on MA-10 LC apoptosis. (A) MA-10 LC were incubated with 10 or 25 µmol/L HEM for 24 h, processed by TUNEL assay and analyzed by flow cytometry. Doxorubicin was used as a positive control for apoptosis. Scale bars are means ± s.e.m. for a representative (n = 3) triplicate experiment. ****P ≤ 0.0001 vs control. (B) Representative immunoblot for caspase-3 protein. The cells were incubated with or without 10 and 25 µmol/L HEM for 24 and 48 h. After treatments, the cells were lysed and subjected to Western blotting analysis.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, TUNEL Assay, Flow Cytometry, Positive Control, Control, Western Blot

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 3 Effect of HEM on MA-10 cell cycle progression. Cells were incubated for 24 h in the absence or presence of 25 µmol/L HEM (A). Then, hemin was withdrawn, and the cells were maintained in fresh GM for another 48 h (B). After the incubation, cells were fixed, permeabilized and stained with PI. DNA content was analyzed by flow cytometry. Graphs show G1/G0, S, and G2/M cell cycle phases distribution. Scale bars are means ± s.e.m. for a representative (n = 3) triplicate experiment. ***P ≤ 0.001.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, Staining, Flow Cytometry

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 4 Effect of long-term exposure to HEM on P4 production in MA-10 LC. The cells were incubated in the presence or absence of 10 µmol/L HEM for 24 or 48 h (A). Then, hemin was withdrawn, and the cells were maintained in fresh growth medium for another 36 or 48 h (B). Progesterone was measured by RIA. Scale bars are means ± s.e.m. for a representative (n = 3) triplicate experiment. **P ≤ 0.01 and ****P ≤ 0.0001 vs control.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, Control

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 5 Interaction between HEM and HA or specific HA receptor agonists in LC steroidogenesis. MA-10 (A and C) and R2C (B and D) cells were preincubated in the presence or absence of 10 µmol/L HEM for 30 min. Then, 1 nmol/L or 10 µmol/L HA or 1 µmol/L FMPH, AMTHA or VUF were added and the incubation continued for 5 h. Progesterone was measured by RIA. Scale bars are means ± s.e.m. for a representative (n = 3) triplicate experiment. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001 vs control.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, Control

Journal: Journal of Molecular Endocrinology

Article Title: Heme oxygenase-1 arrests Leydig cells functions and impairs their regulation by histamine

doi: 10.1530/jme-19-0063

Figure Lengend Snippet: Figure 6 Interaction between HEM and HA or specific HA receptor agonists in LC proliferation. MA-10 (A and C) and R2C (B and D) cells were preincubated in the presence or absence of 25 µmol/L HEM for 30 min. Then, 1 nmol/L or 10 µmol/L HA or 1 µmol/L FMPH, AMTHA or VUF were added and the incubation continued for 24 h. The cells were labeled with [3H]-thymidine during the last 16 h. Scale bars are means ± s.e.m. for a representative (n = 3) octuplicate experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001 vs control.

Article Snippet: The R2C rat Leydig tumor cell line was purchased from ATCC (# 58649146).

Techniques: Incubation, Labeling, Control

Journal: The Journal of Biological Chemistry

Article Title: Plasma Membrane Origin of the Steroidogenic Pool of Cholesterol Used in Hormone-induced Acute Steroid Formation in Leydig Cells *

doi: 10.1074/jbc.M116.740928

Figure Lengend Snippet: Cholesterol trafficking analysis in primary rat Leydig cells infected with lentivirus containing mCherry-D4. A, confocal microscope images of cells with and without addition of Bt2cAMP. Scale bar, 10 μm. B, time course of mCherry-D4 fluorescence intensity at the plasma membrane of control and Bt2cAMP-treated cells. C, testosterone content of control cells and cells infected with lentivirus before and after a 2-h treatment with Bt2cAMP. Testosterone was measured just after the cells were isolated (0 h), in control (uninfected) cells 18 h after incubation and 18 h after infection. Data represent means ± S.D. of at least three independent experiments performed in triplicate; two-way ANOVA followed by Bonferroni's post hoc test (*) was used to calculate statistical significance. *, p < 0.05; **, p < 0.01; ***, p < 0.001. dbcAMP, dibutyryl cAMP.

Article Snippet: MA-10 cells from mouse Leydig cell tumor (kindly provided by Dr. M. Ascoli, University of Iowa, Iowa City) and R2C cells from rat Leydig cell tumor cells (American Type Culture Collection, Manassas, VA) were cultured with Dulbecco's modified Eagle's medium (DMEM)/Ham's F-12 (Invitrogen) supplemented with 1% penicillin and 1% streptomycin and maintained at 37 °C.

Techniques: Infection, Microscopy, Fluorescence, Isolation, Incubation