Journal: Reproductive Sciences

Article Title: The Effect of Thymoquinone on the TNF-α/OTULIN/NF-κB Axis Against Cisplatin-İnduced Testicular Tissue Damage

doi: 10.1007/s43032-024-01567-y

Figure Lengend Snippet: Effect of CS and\or TQ applications on reproductive hormones Serum testosterone and GnRH levels were similar in the control and TQ groups. GnRH levels increased while testosterone levels decreased in the CS group compared to the control group. In the CS + TQ group, it was determined that while testosterone levels increased compared to the CS group, GnRH levels decreased. *; compared to the control group ( p < 0.05), #; Compared with the CS group ( p < 0.05). CS; Cisplatin, TQ; Thymoquinone, GnRH; Gonadotropin-releasing hormone

Article Snippet: ELISA kits were used according to the manufacturer's instructions GnRH (Elabscience, USA), (OTULIN (SunRed, China), MDA, CAT, and SOD kits were purchased from Fine Test (China)).

Techniques: Control

Journal: Reproductive Sciences

Article Title: The Effect of Thymoquinone on the TNF-α/OTULIN/NF-κB Axis Against Cisplatin-İnduced Testicular Tissue Damage

doi: 10.1007/s43032-024-01567-y

Figure Lengend Snippet: Effect of CS and/or TQ applications on OTULIN levels in testicular tissues: A ; OTULIN immunoreactivity microphotographs and graph, B ; OTULIN ELISA levels, C ; OTULIN mRNA expression, D ; OTULIN gene protein level. OTULIN levels in testicular tissues of control and TQ groups were similar. OTULIN immunoreactivity, ELISA level, mRNA expression and gene protein level were decreased in the CS group compared to the control group. An increase in OTULIN levels was detected in the CS + TQ group compared to the CS group. *; compared to the control group ( p < 0.05), #; Compared to CS group ( p < 0.05). A; OTULIN immunohistochemical staining, scale bar; 100 μm. CS; Cisplatin, TQ; Thymoquinone

Article Snippet: ELISA kits were used according to the manufacturer's instructions GnRH (Elabscience, USA), (OTULIN (SunRed, China), MDA, CAT, and SOD kits were purchased from Fine Test (China)).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control, Immunohistochemical staining, Staining

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: Correlation between GnRH expression and malignancy in pancreatic cancer. (A) Oncomine data analysis of GnRH mRNA levels in 16 types of cancer from the Ramaswamy multi-cancer datasets; (B) Representative images of GnRH expression in pancreatic cancer; (C) Pathological analysis of the correlation between GnRH expression and malignancy in pancreatic cancer. (D) Overall survival rates according to data from the TCGA database. ** p <0.01; Scale bars, 50 μm.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: Expressing

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: GnRH regulates pancreatic cancer cell proliferation. (A) the expression levels of GnRH and GnRHR in GnRH-OE, GnRH-KD, or Control group Panc1 cells; (B) Proliferation of GnRH-OE, GnRH-KD, GnRHR-KD, or Control group Panc1 cells; (C) Representative images of colony formation in GnRH-OE, GnRH-KD, or Control group Panc1 cells; (D) Statistical analysis of colony formation in GnRH-OE, GnRH-KD, or Control group Panc1 cells. * p <0.05, ** p <0.01, compared with the control.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: Expressing, Control

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: GnRH treatment inhibits cell proliferation by inducing apoptosis in pancreatic cancer cells. (A) TUNEL assays were performed to determine the number of apoptotic Panc1 cells in the GnRH-OE, GnRH-KD, and Control groups; (B) Percentage of apoptotic Panc1 cells in the GnRH-OE, GnRH-KD, and Control groups; (C,D) Expression levels of Bcl-2, Bax, c-myc, phosphor-c-myc, cleaved caspase-3, and cleaved caspase-9 in GnRH-OE, GnRH-KD, and Control group Panc1 cells. * p <0.05, ** p <0.01, compared with the control; Scale bars, 100 μm.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: TUNEL Assay, Control, Expressing

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: Autophagy is involved in GnRH-mediated apoptosis in pancreatic cancer cells. (A) The expression of Beclin 1, LC3B-I, and LC3B-II was detected via western blot analysis in GnRH-OE, GnRH-KD, and Control group Panc1 cells. Quantitative analysis of the protein Beclin 1 and β-actin expression ratio (B) and LC3-II and LC3-I expression ratio (C) in GnRH-OE, GnRH-KD, and Control group Panc1 cells. * p <0.01, compared with the control. (D,E) LC3 II expression was regulated in CQ- or 3-MA-treated GnRH-OE Panc1 cells. (F) TUNEL assay revealed that 3-MA treatment inhibits apoptosis in GnRH-OE Panc1 cells. (G) Proliferation of GnRH-OE, 3-MA-treated GnRH-OE, or Control group Panc1 cells * p <0.05, ** p <0.01, compared with the control.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: Expressing, Western Blot, Control, TUNEL Assay

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: Effects of GnRH on tumor invasion and migration in pancreatic cancer cells. (A,B) The migration of GnRH-OE, GnRH-KD, or Control group Panc1 cells at 0, 12, 24, and 36 h; (C,D) Cell invasion was determined by transwell assays in GnRH-OE, GnRH-KD, and Control group Panc1 cells at 24 h. (E) MMP2 and MMP9 protein expression was determined in GnRH-OE, GnRH-KD, and Control group Panc1 cells. * p <0.05, ** p <0.01, compared with the control. Scale bar, 200 μm.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: Migration, Control, Expressing

Journal: Frontiers in Endocrinology

Article Title: The Anti-proliferative Activity of GnRH Through Downregulation of the Akt/ERK Pathways in Pancreatic Cancer

doi: 10.3389/fendo.2019.00370

Figure Lengend Snippet: The Akt/ERK pathways are associated with GnRH-mediated cell proliferation through apoptosis induction in pancreatic cancer cells. (A) Expression levels of p-Akt, t-Akt, p-ERK1/2, and t-ERK1/2 were determined by western blotting in GnRH-OE, GnRH-KD, and Control group Panc1 cells; Quantitative analysis of the protein p-Akt and t-Akt expression ratio (B) and p-ERK1/2 and t-ERK1/2 expression ratio (C) in GnRH-OE, GnRH-KD, and Control group Panc1 cells; (D) Proliferation of GnRH-inhibited Panc1 cells with or without MK-2206 or SCH772984 treatment; Representative images (E) and quantitative analysis (F) of apoptosis of GnRH-inhibited Panc1 cells with or without MK-2206 or SCH772984 treatment. p-Akt, phosphorylated-Akt; t-Akt, total-Akt; p-ERK1/2, phosphorylated-ERK1/2; t-ERK1/2, total-ERK1/2. * p <0.05, ** p <0.01, compared with the control. Scale bars, 100 μm.

Article Snippet: The GnRH Crispr Activation Plasmids system, including GnRH Crispr Activation Plasmid, the Crispr/dCas9-VP64-Blast plasmid, and MS2-P65-HSF1-Hygro plasmid (sc-401425-ACT, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the GnRH HDR Plasmid (h2) system, including GnRH HDR Plasmid (h2), and GnRH Crispr/Cas9 KO plasmid (sc-401425-HDR-2, Santa Cruz Biotechnology), were used to overexpress or knockdown GnRH expression in pancreatic cancer cells, respectively.

Techniques: Expressing, Western Blot, Control

Journal: Heliyon

Article Title: Brucella abortus RB51 Δ leuB expressing Salmonella FliC conjugated gonadotropins reduces mouse fetal numbers: A possible feral swine brucellosis immunocontraceptive vaccine

doi: 10.1016/j.heliyon.2021.e06149

Figure Lengend Snippet: Anti-his-tag Western blot. Fluorescent anti-his-tag antibodies were applied to blotted protein extracts. Lane-1 consists of the BioRad Precision Plus Protein Ladder. Lane-2 consists of protein extract from strain RB51L, showing no significant bands. Lane-3 consists of protein extract from strain RB51LGnRH isolated from the spleens of BABLB/c mice 14 days post inoculation, with a his-tag protein between 37 kDa and 50 kDa. This corresponds with the theoretical size of the his-tagged FliC-GnRH protein (≈40kDa). The original pictograph is represented in Supplemental Figure 4 .

Article Snippet: The presence of GnRH and FSH antibodies was confirmed via absorbance values with the following protocol: Antigens (Provided by Boster and BEI Resources, VA, USA) were diluted in PBS to a concentration of 20 μg/ml.

Techniques: Western Blot, Isolation

Journal: Heliyon

Article Title: Brucella abortus RB51 Δ leuB expressing Salmonella FliC conjugated gonadotropins reduces mouse fetal numbers: A possible feral swine brucellosis immunocontraceptive vaccine

doi: 10.1016/j.heliyon.2021.e06149

Figure Lengend Snippet: GnRH and porcineFSHβ IgG antibody ELISAs from male and female BALB/c mice. (a) Fifty microliters of serum collected 81 days post vaccination from both male and female mice were tested in duplicate for detection of anti-GnRH IgG antibodies. All group values were compared via One-way ANOVA, and the statistically significant differences are shown. The OD 450 values from the serum of mice in the strain RB51LGnRH immunized group were significantly higher than those from the PBS control group with a p value of <0.0001. The OD 450 values from the serum of mice in the Improvest® immunized group were significantly higher than those from the control group PBS, with the p value of <0.0001. The OD 450 values from the serum of mice in the strain RB51LGnRH immunized group were significantly higher than those from the RB51L control group with a p value of 0.02. (b) Fifty microliters of serum collected 81 days post vaccination from both male and female mice were tested in duplicate for detection of porcineFSHβ IgG antibodies. All group values were compared via One-way ANOVA, and the statistically significant differences are shown. The OD 450 values from the serum of mice in the strain RB51LF immunized group were significantly higher than those from the PBS control group with a p value of <0.0001 and the control RB51L group with a p value of 0.0011. This shows that there was an immune response amounted to the vaccines, which can be further seen by the other data sets. There were no significant differences between groups without brackets and p values.

Article Snippet: The presence of GnRH and FSH antibodies was confirmed via absorbance values with the following protocol: Antigens (Provided by Boster and BEI Resources, VA, USA) were diluted in PBS to a concentration of 20 μg/ml.

Techniques: Control, Vaccines