anova or t-students test calculations using graphpad prism v.9.4.1  (GraphPad Software Inc)

Journal: The Journal of Biological Chemistry

Article Title: AMH regulates a mosaic population of AMHR2-positive cells in the ovarian surface epithelium

doi: 10.1016/j.jbc.2024.107897

Figure Lengend Snippet: Granulosa factor(s) suppress MOSE cell growth in culture. A , primary MOSE and OVD cells were isolated from wild-type mice and expanded in culture. Proliferation of cells was determined using a WST-1 assay, where cell number correlates with the absorbance measured at 450 nm. Relative cell numbers were measured in triplicate samples and are represented as the mean ± s.d. B , both MOSE and OVD have a characteristic cuboidal epithelial shape in culture. C , MOSE cells were co-cultured in transwell dishes with (+G) or without (control) granulosa cells (+G). Cell number was determined by the WST-1 proliferation assay kit. D , Conditioned medium was prepared from granulosa cells after 2 (2 days) or 4 (4 days) days in culture. MOSE cells were plated in triplicate in 96-well dishes and grown overnight, then incubated in granulosa cell conditioned medium (GCCM) mixed with normal culture medium at the percentages shown; cell number was assayed after 4 days in culture, when MOSE cells are actively proliferating. Data are expressed as mean ± s.d., with ∗ p < 0.05, by Student’s unpaired two-tailed t test. E , representative images of MOSE cells co-cultured without (Control) or with (+Granulosa) granulosa cells.

Article Snippet: The Student’s unpaired two-tailed t test calculator offered online from GraphPad was used to compare and test for significance between two groups, where p < 0.05 is considered statistically significant.

Techniques: Isolation, WST-1 Assay, Cell Culture, Control, Proliferation Assay, Incubation, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: AMH regulates a mosaic population of AMHR2-positive cells in the ovarian surface epithelium

doi: 10.1016/j.jbc.2024.107897

Figure Lengend Snippet: AMH suppresses MOSE cell growth. A , MOSE cells were treated with TGF-β (0.1 μg/ml) or AMH (5 μg/ml) for 4 days, and cell growth was determined by WST-1 assay and expressed as the cell number of treated cells relative to control cells (fraction of control). The average of three experiments is shown as mean ± s.d., with ∗ p < 0.05 or ∗∗ p = 0.001, determined by an unpaired two-tailed Student’s t test. B , images of cells treated with TGF-β or AMH, as described.

Article Snippet: The Student’s unpaired two-tailed t test calculator offered online from GraphPad was used to compare and test for significance between two groups, where p < 0.05 is considered statistically significant.

Techniques: WST-1 Assay, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: AMH regulates a mosaic population of AMHR2-positive cells in the ovarian surface epithelium

doi: 10.1016/j.jbc.2024.107897

Figure Lengend Snippet: AMH from granulosa cell conditioned medium suppresses MOSE growth in culture. A , AMH was immunodepleted from granulosa cell conditioned medium (GCCM) using an excess concentration of anti-AMH rabbit polyclonal antibody (Abcam ab84952) (Anti-AMH). Controls included normal culture medium (control) and granulosa cell conditioned medium that had no treatment (none), treated only with the affinity matrix used for IP (+ protein A/G), or non-specific pre-immune rabbit serum and Protein A/G (Pre-Ab). MOSE cultures were treated with the respective medium for 6 days, and relative cell number was determined by WST-1 assay, shown as mean ± s.d., with ∗ p < 0.05, as determined by an unpaired two-tailed Student’s t test. The error bars indicate multiple and potentially overlapping measurements. There was no significant difference (N.S.) between cell proliferation in control medium and anti-AMH immunodepleted granulosa cell conditioned medium; however, a significant difference was determined between cells grown in untreated granulosa cell conditioned medium (none) and anti-AMH immunodepleted granulosa cell conditioned medium. B , granulosa cells were isolated from wild-type or AMH knockout mice and grown in culture for 6 days. The conditioned medium was collected and incubated with MOSE cells for an additional 6 days. The effect of granulosa cell conditioned medium from normal wild type or AMH knockout mouse ovaries was compared to normal culture medium. MOSE cell proliferation was significantly decreased by growth in WT GCCM, determined as ∗ p < 0.05 by an unpaired two-tailed Student’s t test. C , MOSE cells were treated with TGFβ, EGF, or granulosa cell conditioned medium for 6 days, then collected in sample buffer, and analyzed by Western blot for the proteins indicated. D , images of cells treated with TGFβ, EGF, and conditioned medium collected from wild type (WT) or AMH KO granulosa cells grown in culture for 6 days.

Article Snippet: The Student’s unpaired two-tailed t test calculator offered online from GraphPad was used to compare and test for significance between two groups, where p < 0.05 is considered statistically significant.

Techniques: Concentration Assay, Control, WST-1 Assay, Two Tailed Test, Isolation, Knock-Out, Incubation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: AMH regulates a mosaic population of AMHR2-positive cells in the ovarian surface epithelium

doi: 10.1016/j.jbc.2024.107897

Figure Lengend Snippet: Expansion of AMHR2-Cre+;Rosa ± MOSE and OVD epithelial cells in vitro . A–B , primary cultures of MOSE and (C-D) OVD epithelial cells were established from 4-month-old AMHR2-Cre+;Rosa ± mice and allowed to expand in culture for 1 to 4 weeks. LacZ (X-Gal) staining of the cultures was performed. B – D , the percentage of X-Gal positive cells was calculated from an average of at least three separate fields, expressed as mean ± s.d. Significant difference from the starting time point was calculated for each subsequent time point using an unpaired two-tailed Student’s t test, shown as ∗ p < 0.05 and ∗∗ p = 0.0001. Results are representative of three separate experiments. Images were taken at 63 × magnification.

Article Snippet: The Student’s unpaired two-tailed t test calculator offered online from GraphPad was used to compare and test for significance between two groups, where p < 0.05 is considered statistically significant.

Techniques: In Vitro, Staining, Two Tailed Test