Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Effects of cordycepin on cell proliferation and morphology on FGF9-treated TM3 cells. MTT assay for the viability of FGF9-treated TM3 cells. After 18 h of serum starvation, TM3 cells were co-treated with or without 50 ng/ml FGF9 and 0, 25, 50, 100 μM cordycepin for 12 (A) and 24 h (B). Examination of morphology and cell growth were observed under light microscopy and cells were co-treated with or without 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 (C) and 24 h (D). All values are represented as the mean±standard error of the mean (SEM), n=3. p-Values were calculated using one-way ANOVA followed by Tukey’s multiple comparisons test. Different letters above the bars represent statistical differences among different treatments; p<0.05. Yellow arrowheads in (C) and (D) indicate apoptotic cells with rounded up phenomena.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: MTT Assay, Light Microscopy

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Effects of cordycepin on colony formation on FGF9-treated TM3 cells. After co-treatment with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 of μM cordycepin for 12 and 24 h, TM3 cells (A and B) were trypsinized and collected. 1,500 cells were then replated in 6-cm culture dishes to allow for colony development. After 8-14 days, colonies were stained by 0.5 % crystal violet solution. The number of colonies between different treatments were then counted and were analyzed by two-way ANOVA with Tukey’s multiple comparisons test (C and D). All values are represented as the mean±standard error of the mean (SEM), n=3 for 12 and 24 h. *p<0.05, **p<0.01 and ***p<0.001 vs. control group, ##p<0.01 and ###p<0.001 vs. the FGF9 alone group, $p<0.05 and $$p<0.01 vs. combined group.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Staining, Control

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin did not induce cell apoptosis in FGF9-treated TM3 cells. After co-treatment with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin in TM3 cells for 12 (A) and 24 h (B), cells were stained with annexin V and propidium iodide (PI) and analyzed by flow cytometry. All values are represented as the mean±standard error of the mean (SEM), n=3. Two-way ANOVA with Tukey’s multiple comparisons test was used to compare differences between different groups for 12 (C) and 24 h (D). There are no statistical differences between treatments in (C) and (D).

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Staining, Flow Cytometry

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin did not induce apoptosis-related protein expression on FGF9-treated TM3 cells. Western blot analysis for the expression of cleaved caspase-8, cleaved caspase-9, cleaved caspase-3 and cleaved PARP in TM3 cells co-treated with or without 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 (A) and 24 (B) h. All values are represented as the mean±standard error of the mean (SEM), n=3. Two-way ANOVA with Tukey’s multiple comparisons test was used to compare differences between different groups. P, positive control.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Expressing, Western Blot, Positive Control

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin induced autophagy in FGF9-treated TM3 cells. After co-treatment with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 and 24 h, TM3 cells were stained with Acridine orange (AO) for acidic vesicular organelles (AVOs) through flow cytometric analysis and analyzed (A and B). Quantification and analysis of histograms upon autophagy cells from flow cytometric analysis are illustrated at 12 (C) and 24 h (D). All values are represented as the mean±standard error of the mean (SEM), n=3. Two-way ANOVA with Tukey’s multiple comparisons test were used to compare differences between different groups. Different letters above bars represent statistical differences among different treatments; p<0.05.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Staining

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin induced autophagy in FGF9-treated TM3 cells. After co-treatment with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 and 24 h, LC3 puncta were detected by immunofluorescent staining. Images were analyzed for bright objects using the analyze particles tool in ImageJ (A and B). Histograms of quantification of autophagic cells upon LC3 puncta immunofluorescent staining are illustrated at 12 (A) and 24 h (B). All values are represented as the mean±standard error of the mean (SEM), n=3. Two-way ANOVA with Tukey’s multiple comparisons test were used to compare differences between different groups. Different letters above bars represent statistical differences among different treatments; p<0.05. Scale bars are equal to 10 μm.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Staining

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin didn’t induce the expression of Atg5-12 on FGF9-treated TM3 cells. TM3 cells were co-treated with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 and 24 h, respectively. Western blot analyses for the expression of Atg5-12 for 12 and 24 h, in TM3 cells were examined (A and B). Results in (C) and (D) for 12- and 24-h treatments are shown as mean±standard error of the mean (SEM) with at least 3 independent experiments. Two-way ANOVA with Tukey’s multiple comparisons tests was used to determine differences among different treatments. There are no statistical differences between treatments in (C) and (D).

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin did not induce the expression of beclin-1 on FGF9-treated TM3 cells. TM3 cells were co-treated with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 and 24 h. Western blot analyses for the expression of beclin-1 for 12 and 24 h in TM3 cells were examined (A and B). Results in (C) and (D) for 12- and 24-h treatments are shown as mean±standard error of the mean (SEM) with at least 3 independent experiments. Two-way ANOVA with Tukey’s multiple comparisons tests was used to determine differences among different treatments. There are no statistical differences between treatments in (C) and (D).

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Cordycepin induced the expression of LC3-I/II on FGF9-treated TM3 cells. TM3 cells were co-treated with 0 or 50 ng/ml FGF9 plus 0, 25, 50, and 100 μM cordycepin for 12 and 24 h. Western blot analyses for the expression of LC3-I/II for 12 and 24 h in TM3 cells were examined (A and B). Results in (C) and (D) for 12- and 24-h treatments are shown as mean±standard error of the mean (SEM) with at least 3 independent experiments. Two-way ANOVA with Tukey’s multiple comparisons tests were used to determine differences among different treatments. Different alphabetical letters above bars represent statistical differences among different treatments; p<0.05.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Cancer Genomics & Proteomics

Article Title: Cordycepin Activates Autophagy to Suppress FGF9-induced TM3 Mouse Leydig Progenitor Cell Proliferation

doi: 10.21873/cgp.20479

Figure Lengend Snippet: Autophagy inhibitor rescued cordycepin-induced cell death on FGF9-treated TM3 cells. TM3 cells (6.5×103 cells per well) were seeded in 96 well plates with 100 μl of culture medium and pretreated with 100 μM CQ (autophagy inhibitor) for 1 h. Then, TM3 cells were co-treated with or without 50 ng/ml FGF9 and 50 μM cordycepin for 24 h. The PrestoBlue assay was then used to assess cell viability. All values are represented as the mean±standard error of the mean (SEM), n=3. Two-way ANOVA with Tukey’s multiple comparisons test was used to compare differences between different groups. Different letters above bars represent statistical differences among different treatments; p<0.05.

Article Snippet: Human fibroblast growth factor 9 (FGF9) was procured from PeproTech (Rocky Hill, NJ, USA).

Techniques: Prestoblue Assay