Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Cellular viability was correlated to Nur77 expression induced by venlafaxine. (A) Chemical structure of venlafaxine. (B) Dose-dependent inhibition of venlafaxine on the growth inhibition of MV3 cells at 72 h. (C) Relative mRNA expressions of Nur77 in MV3 cells were quantified after treatment with venlafaxine (10 μM) for 0–8 h. (D) The protein expression of Nur77 in MV3 cells was detected by Western blot after treatment with venlafaxine (10 μM) for 0–8 h. Intensity of the protein bands was quantified and normalized to loading control GAPDH. N = 3, ***, p < .001 vs. control (0 h).

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Expressing, Inhibition, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Venlafaxine induced apoptosis in MV3 cells. (A) The protein expression of BAX-2, cleaved caspase 3 and cleaved PARP in MV3 cells was detected by Western blot after treatment with venlafaxine (10 μM) for 0–8 h. Intensity of the protein bands was quantified and normalized to loading control GAPDH. (B) Apoptosis assays in MV3 cells treated with venlafaxine (10 μM) for 0–8 h were conducted by flow cytometry. (C) The apoptotic cells were detected by TUNEL assay in MV3 cells treated with venlafaxine (10 μM) for 0–8 h. N = 3, ***, p < .001 vs. control (0 h).

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Expressing, Western Blot, Control, Flow Cytometry, TUNEL Assay

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Venlafaxine induced Nur77 mitochondrial targeting and ROS production in MV3 cells. (A) MV3 cells treated with venlafaxine (10 μM) for 0–8 h were immunostained with Bcl-2 and Nur77 antibodies and visualized by confocal microscopy. (B) MV3 cells treated with the cellular ROS assay kit and visualized by confocal microscopy.

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Confocal Microscopy, ROS Assay

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Nur77 expression was necessary for venlafaxine-induced apoptosis. MV3 cells were transfected with siRNA control or Nur77 siRNA, following by treatment with venlafaxine (10 μM) for 0–8 h. (A) The protein expression of BAX-2, caspase 3 and cleaved PARP in MV3 cells was detected by western blot. Inte cleaved nsity of the protein bands was quantified and normalized to loading control GAPDH. (B) Apoptosis assays in MV3 cells were conducted by flow cytometry. (C) The apoptotic cells were detected by TUNEL assay. (D) MV3 cells treated with the cellular ROS assay kit and visualized by confocal microscopy. N = 3, **, p < .01, ***, p < .001 vs. control.

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Expressing, Transfection, Control, Western Blot, Flow Cytometry, TUNEL Assay, ROS Assay, Confocal Microscopy

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Activation of ERK1/2 signaling was necessary for venlafaxine-induced apoptosis. (A) Representative western-blot bands and quantification of the MAPK signal molecules abundances in MV3 cells treated with venlafaxine (10 μM) for 0–8 h (B) MV3 cells were treated with vehicle, JNK1/2 inhibitor SP600125 and MERK/ERK inhibitor PD98059 for 72 h. Cell viability were measured by CCK-8 kits at 72 h (C) MV3 cells were transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 72 h. Cell viability were measured by CCK-8 kits at 72 h (D,E) Representative western-blot bands and quantification of Nur77 abundances in MV3 cells treated with venlafaxine (10 μM) for 8 h, or transfected with siRNA control, JNK1/2 siRNA, and ERK siRNA for 20 h (F) MV3 cells were immunostained with Bcl-2 and Nur77 antibodies and visualized by confocal microscopy. N = 3, *, p < .05, **, < .01, ***, p < .001 vs. control.

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Activation Assay, Western Blot, CCK-8 Assay, Transfection, Control, Confocal Microscopy

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Anti-cancer efficacy of venlafaxine in mice. BALB/c nude mice bearing MV3 or Nur77 KO MV3 xenograft tumors were treated with venlafaxine (20 mg/kg, i.p.) or its vehicle once daily from week 1 to week 5 after implantation of MV3 or Nur77 KO MV3 cells. (A) Tumor tissues were immunostained with Nur77 and JNK1/2 antibodies and visualized by confocal microscopy. (B) Venlafaxine reduced cancer cell growth in nude mice through Nur77. One of five similar experiments is shown.

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: Confocal Microscopy

Journal: Frontiers in Pharmacology

Article Title: Venlafaxine, an anti-depressant drug, induces apoptosis in MV3 human melanoma cells through JNK1/2-Nur77 signaling pathway

doi: 10.3389/fphar.2022.1080412

Figure Lengend Snippet: Venlafaxine-induced apoptosis in vivo . BALB/c nude mice bearing WT MV3 or Nur77 KO MV3 xenograft tumors were treated with venlafaxine (20 mg/kg, i.p.) or its vehicle once daily from week 1 to week 5 after implantation of MV3 or Nur77 KO MV3 cells. (A) Representative western-blot bands and quantification of BAX-2, cleaved caspase 3 and cleaved PARP abundances in tumor tissues. N = 5, *, p < .05, **, < .01, ***, p < .001 vs. MV3 vehicle control. ### , p < .001 vs. MV3 venlafaxine control. (B) Tumor tissues were immunostained with cleaved caspase 3 and cleaved PARP antibodies and visualized by confocal microscopy. (C) Tumor tissues were immunostained with TUNEL assay kits and antibodies and Ki-67 visualized by confocal microscopy. One of five similar experiments is shown.

Article Snippet: Human melanoma (MV3) cells were purchased from the China center for type culture collection (Cat. #GDC0615).

Techniques: In Vivo, Western Blot, Control, Confocal Microscopy, TUNEL Assay

Journal: Yonsei Medical Journal

Article Title: Efficiency of Recombinant Bacille Calmette-Guérin in Inducing Humoral and Cell Mediated Immunities against Human Immunodeficiency Virus Type 1 Third Variable Domain in Immunized Mice

doi: 10.3349/ymj.2011.52.1.173

Figure Lengend Snippet: (A) V3-trimer was cloned into pRSET-B vector, and expressed in E. coli BL21 (DE3). Whole cell lysates or purified protein was separated on a SDA-PAGE. Lane 1, pRSET/B control vector-transformed cell lysate; lane 2, pRSET-mV3-transformed cell lysate; lane 3, Ni + -NTA resin-purified recombinant mV3 protein. (B) Western blot analysis of V3-concatamer (three V3) expressed in bacteria with 2 different antisera. pRSET/B control vector-transformed cell lysate (1) and pRSET-mV3-transformed cell lysate (2) were separated on a SDS-PAGE, and then assessed by Western blot analysis with mouse anti-3V3-antibody (a) and rabbit anti-gp120 antibody (b). SDA-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; V3, third variable.

Article Snippet: Analysis of mV3-specific antibodies was performed with a mouse Ig isotyping kit (Pharmingen, San Diego, CA, USA).

Techniques: Clone Assay, Plasmid Preparation, Purification, Transformation Assay, Recombinant, Western Blot, SDS Page, Polyacrylamide Gel Electrophoresis

Journal: Yonsei Medical Journal

Article Title: Efficiency of Recombinant Bacille Calmette-Guérin in Inducing Humoral and Cell Mediated Immunities against Human Immunodeficiency Virus Type 1 Third Variable Domain in Immunized Mice

doi: 10.3349/ymj.2011.52.1.173

Figure Lengend Snippet: (A) Western blot analysis of rBCG-mV3 lysates with anti-V3 antiserum. Cultures of rBCG-mV3 were heat-induced at 45 for 2 hours in the log phase, and the same amounts of cells were harvested from the culture in 2 week intervals. M, marker; lane 1, BCG-pMV control; lane 2, BCG-mV3 (2 weeks); lane 3, BCG-mV3 (4 weeks); lane 4, BCG-mV3 (6 week) after a single heat-shock; lane 5, mV3-expressing E. coli lysate as a positive control. (B) PCR amplification of the plasmid extracted from the kanamycin-resistant rBCG that was isolated from the spleen of immunized mice was performed with V3-specific primer set. Each lane shows a pMV-mV3 positive control (1), pMV261 negative control (2), and plasmid recovered from the rBCG-mV3-immunized mice 4 weeks (3) and 8 weeks after immunization (4). (C) Western blot analysis of rBCG-mV3 recovered from immunized mice with anti-mV3 sera. Each lane shows BCG-pMV (1), rBCG-mV3 (2), and pRSET-mV3 / E. coli BL21 (DE3) lysate (3). BCG, Bacille Calmette-Guérin; PCR, polymerase chain reaction.

Article Snippet: Analysis of mV3-specific antibodies was performed with a mouse Ig isotyping kit (Pharmingen, San Diego, CA, USA).

Techniques: Western Blot, Marker, Expressing, Positive Control, Amplification, Plasmid Preparation, Isolation, Negative Control, Polymerase Chain Reaction

Journal: Yonsei Medical Journal

Article Title: Efficiency of Recombinant Bacille Calmette-Guérin in Inducing Humoral and Cell Mediated Immunities against Human Immunodeficiency Virus Type 1 Third Variable Domain in Immunized Mice

doi: 10.3349/ymj.2011.52.1.173

Figure Lengend Snippet: rBCG-mV3 induces V3-specific antibody in BALB/c mice. (A) Six-week old BALB/c mice were inoculated with control (transformed with pMV261 empty vector) and recombinant BCG (rBCG-mV3) at a concentration of 1 × 10 7 cells/mouse intraperitoneally. The titer of anti-V3 antibody in the serum obtained from the immunized mice every 2 weeks were assessed by ELISA as described in the Materials and Methods. Anti-serum was collected from the rBCG-mV3-immunized mice 6 weeks after primary inoculation. (B) Cell lysates or purified mV3 were examined by Western blot analysis with the antiserum obtained from the mice immunized with rBCG-mV3. The arrow indicates the band of purified mV3 protein. (C) C8166 human T cell line was infected with 1 MOI of HIV-1 (HXB2), which was pre-incubated with mouse anti-rBCG-mV3 sera or control anti-BCG sera at 37 for 1 hr. Cells were harvested and lyzed at the indicated time point and then p24 antigen was assessed by Western blot analysis with anti-p24 monoclonal antibody. OD, optical density; PI, postinoculation; MOI, multiplicities of infection; BCG, Bacille Calmette-Guérin.

Article Snippet: Analysis of mV3-specific antibodies was performed with a mouse Ig isotyping kit (Pharmingen, San Diego, CA, USA).

Techniques: Transformation Assay, Plasmid Preparation, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay, Purification, Western Blot, Infection, Incubation

Journal: Yonsei Medical Journal

Article Title: Efficiency of Recombinant Bacille Calmette-Guérin in Inducing Humoral and Cell Mediated Immunities against Human Immunodeficiency Virus Type 1 Third Variable Domain in Immunized Mice

doi: 10.3349/ymj.2011.52.1.173

Figure Lengend Snippet: (A) Lymphocyte proliferation assay in response to recombinant-mV3 antigen stimulation in T cell-enriched splenocytes of immunized mice. Spleens were removed from 3 mice per group 10 weeks and 5 months after immunization with rBCG-mV3 and control BCG (BCG-PMV), respectively. Lymphocyte proliferation assay was performed by measuring the [ 3 H]-thymidine incorporation, and is expressed as the mean SD stimulation indices in triplicate and three independent experiments. (B) Anti-mV3 antibody isotyping was performed using a monoclonal antibody-based mouse Ig isotyping kit and expressed by percentage for each isotype and subclass. Anti-serum was obtained from mice 10 weeks after immunization with rBCG-mV3. Anti-serum was pre-cleared with BCG lysates and then used for ELISA. PI, postinoculation; BCG, Bacille Calmette-Guérin.

Article Snippet: Analysis of mV3-specific antibodies was performed with a mouse Ig isotyping kit (Pharmingen, San Diego, CA, USA).

Techniques: Lymphocyte Proliferation Assay, Recombinant, Enzyme-linked Immunosorbent Assay