Journal: International journal of molecular sciences

Article Title: GABBR2 as a Downstream Effector of the Androgen Receptor Induces Cisplatin Resistance in Bladder Cancer.

doi: 10.3390/ijms241813733

Figure Lengend Snippet: Figure 4. Effects of GABBR2 inhibitor or knockdown on the viability of bladder cancer cells. The MTT assay in UMUC3 (A), 5637-AR (B), or 647V-AR (C) cultured in the absence or presence of various doses of CGP46381 or UMUC3-control-shRNA vs. UMUC3-GABBR2-shRNA (D) for 48 h. Cell viability representing the mean (+SD) from a total of six determinants is presented relative to that of mock treatment or control subline.

Article Snippet: DHT and HF, CGP46381, and CDDP were obtained from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher (Waltham, MA, USA), and Santa Cruz Biotechnology (Dallas, TX, USA), respectively.

Techniques: Knockdown, MTT Assay, Cell Culture, Control, shRNA

Journal: International journal of molecular sciences

Article Title: GABBR2 as a Downstream Effector of the Androgen Receptor Induces Cisplatin Resistance in Bladder Cancer.

doi: 10.3390/ijms241813733

Figure Lengend Snippet: Figure 5. Effects of GABBR2 inhibitor or knockdown on the migration of bladder cancer cells. The wound-healing assay in UMUC3 (A), 5637-AR (B), or 647V-AR (C) without or with CGP46381 (5 µM) or UMUC3-control-shRNA vs. UMUC3-GABBR2-shRNA (D) cultured for 24 h after scratching. Cell migration determined by the rate of cells filling the wound area is presented relative to that of mock treatment or control subline. Each value represents the mean (+SD) from a total of six determinants.

Article Snippet: DHT and HF, CGP46381, and CDDP were obtained from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher (Waltham, MA, USA), and Santa Cruz Biotechnology (Dallas, TX, USA), respectively.

Techniques: Knockdown, Migration, Wound Healing Assay, Control, shRNA, Cell Culture

Journal: International journal of molecular sciences

Article Title: GABBR2 as a Downstream Effector of the Androgen Receptor Induces Cisplatin Resistance in Bladder Cancer.

doi: 10.3390/ijms241813733

Figure Lengend Snippet: Figure 7. Effects of GABBR2 inhibitor on CDDP cytotoxicity in bladder cancer cells. The MTT assay in UMUC3 (A), 5637-AR (B), or 647V-AR (C) without or with CGP46381 (5 µM) or UMUC3-control- shRNA or UMUC3-GABBR2-shRNA (D) cultured for 48 h in the absence or presence of CDDP (5 µM). Cell viability representing the mean (+SD) from a total of six determinants is presented relative to that of mock or CGP46381 treatment (without vs. with CDDP) in each subline. * p < 0.05 [vs. CDDP(+)/CGP46381(−) cells].

Article Snippet: DHT and HF, CGP46381, and CDDP were obtained from Sigma-Aldrich (St. Louis, MO, USA), Thermo Fisher (Waltham, MA, USA), and Santa Cruz Biotechnology (Dallas, TX, USA), respectively.

Techniques: MTT Assay, Control, shRNA, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Gamma-Aminobutyric Acid (GABA) Promotes Growth in Zebrafish Larvae by Inducing IGF-1 Expression via GABA A and GABA B Receptors

doi: 10.3390/ijms222011254

Figure Lengend Snippet: GABA A and GABA B receptors are responsible for GABA-mediated growth promotion in zebrafish larvae. Zebrafish larvae ( n = 20) at three days post-fertilization (dpf) were pretreated with 10 µM each of ( A ) bicuculline (GABA A antagonist), ( B ) CGP 46381 (GABA B antagonist), and ( C ) TPMPA (GABA C antagonist) 2 h before treatment with 25 mM GABA. The media were replaced at 6 dpf with GABA, and total body length was measured at 9 dpf. Graphs represent the total body length ( bottom ). Significant differences among the groups were determined using Student’s t -test. All data are presented as mean ± standard error of the mean (** p < 0.01 and *** p < 0.001). N.S., non-significant.

Article Snippet: Bicuculline, CGP 46381, and TPMPA were purchased from Tocris Bioscience (Bristol, United Kingdom).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Gamma-Aminobutyric Acid (GABA) Promotes Growth in Zebrafish Larvae by Inducing IGF-1 Expression via GABA A and GABA B Receptors

doi: 10.3390/ijms222011254

Figure Lengend Snippet: GABA upregulates growth-stimulating gene expression via GABA A and GABA B receptors. Zebrafish larvae ( n = 20) at three days post-fertilization (dpf) were treated with 10 µM of each ( A ) bicuculline, ( B ) CGP 46381, and ( C ) TPMPA 2 h before treatment with 25 mM GABA, and total RNA was extracted at 9 dpf. The RNA was reverse-transcribed, and synthetic cDNA was amplified to determine expression of the genes for insulin-like growth factor 1 ( zIGF-1) , growth hormone 1 ( zGH-1 ), growth hormone receptor 1 ( zGHR-1 ), and cholecystokinin A ( zCCKA ). z β-Actin was used as an internal control. Densitometry analysis was conducted to determine the expression level of each gene relative to that of β-actin and illustrated ( bottom ). Significant differences among the groups were determined using Student’s t -test. All data are presented as mean ± standard error of the mean (*** p < 0.001). N.S., non-significant.

Article Snippet: Bicuculline, CGP 46381, and TPMPA were purchased from Tocris Bioscience (Bristol, United Kingdom).

Techniques: Gene Expression, Reverse Transcription, Amplification, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Gamma-Aminobutyric Acid (GABA) Promotes Growth in Zebrafish Larvae by Inducing IGF-1 Expression via GABA A and GABA B Receptors

doi: 10.3390/ijms222011254

Figure Lengend Snippet: Inhibition of GABA A and GABA B receptors downregulates GABA-mediated IGF-1 and IGF-1R expression in preosteoblast MC3T3-E1 cells. MC3T3-E1 cells (1 × 10 4 cells/mL) were treated with 5 µM each of ( A , D ) bicuculline, ( B , E ) CGP 46381, and ( C , F ) TPMPA 2 h before treatment with 10 mM GABA for seven days. Total RNA was extracted, and RT-PCR was performed to determine the expression level of IGF-1 and IGF-1R . Glyceraldehyde 3-phosphate dehydrogenase gene ( GAPDH ) was used as an internal control. The expression of IGF-1 and IGF-1R relative to that of GAPDH level was determined ( bottom ). In a parallel experiment, cell-free supernatants were collected, and ELISA was conducted. Significant differences among the groups were determined using Student’s t -test. All data are presented as mean ± standard error of the mean (*** p < 0.001). N.S., non-significant.

Article Snippet: Bicuculline, CGP 46381, and TPMPA were purchased from Tocris Bioscience (Bristol, United Kingdom).

Techniques: Inhibition, Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay