Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 1. Effect of androgen on MALT1 expression in AR-positive prostate carcer cells. The LNCaP cells were treated with various concentrations of R1881 for 24 h. The cells were lysed and then MALT1,

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 2. Effects of androgen and MALT1 on PSA expression in androgen-dependent LNCaP prostate cancer cells. The mock-knockdowned LNCaP (LN_shCOL) and MALT1-knockdowned LNCaP (LN_shMALT1) cells were treated with various concentrations of R1881 as indicated for 24 h. The cells were lysed, then MALT1, PSA, and β-actin were determined by (A) immunoblot assays and (B) quantitative analysis. (C) The reporter activity of the PSA reporter vector was cotransfected with various concentrations of MALT1 expression vectors in LNCaP cells. (D) The reporter activity of the PSA reporter vector was cotransfected with MALT1 and/or AR expression vectors and treated with/without 10 nM R1881 in PCJ cells. (E) The reporter activity of the PSA reporter vector was cotransfected with MALT1 expression vectors (white bars) or control vectors (pcDNA3, black bars), and then treated with various concentrations of R1881 as indicated for 24 h in the LNCaP cells. The data are presented as mean percentages (±SE, n = 6) relative to the control vehicle-treated group. (F) The luciferase activity of the PSA reporter vector was treated with/without 10 nM R1881 and 0.1 µM MDV3100 as indicated for 24 h in LNCaP or transient overexpressed AR PCJ cells. * p < 0.05, ** p < 0.01.

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Expressing, Western Blot, Activity Assay, Plasmid Preparation, Control, Luciferase

Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 3. Effect of androgen on NF-κB signaling is dependent on MALT1 in androgen-dependent LNCaP prostate cancer cells. The LN_shCOL and LN_shMALT1 cells were treated with or without 10 nM of R1881 for 24 h. (A) The expression of GAPDH in cytosol and Lamin B1 in the nucleus were determined by immunoblotting after the separation of nuclear and cytoplasmic fractions as indicated. The expression of MALT1, IκBα, phospho-IκBα, and GAPDH in the cytosol fraction (B), as well as p65, p50, and Lamin B1 in the nuclear fraction (C), were determined by immunoblotting. The numbers indicate the ratio of target proteins/GAPDH or Lamin B1 in relation to vehicle treatment. (D) The NF-κB (p65)-binding activity in the LN_shCOL and LN_shMALT1 cells after treatment with R1881 for 18 h. (E) The reporter activity of the MALT1 reporter vector was treated with various concentrations of R1881 as indicated for 24 h in the 22Rv1 cells. The data are presented as mean percentages (±SE, n = 6) relative to the control vehicle-treated group. ** p < 0.01, NS represents no significance.

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Expressing, Western Blot, Binding Assay, Activity Assay, Plasmid Preparation, Control

Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 4. Androgen does not affect MALT1 expression in PC-3 cells and ectopic ARFL-overexpressed PC-3 cells. (A) The PC-DNA cells were treated with various concentrations of R1881 as indicated for 24 h. The cells were lysed, and the MALT1 and β-actin expression was determined using immunoblot assays. The numbers indicate the ratio of target proteins/β-actin in relation to vehicle-treatment. (B) The PC-ARFL cells were treated with various concentrations of R1881 as indicated for 24 h. The cells were lysed and analyzed for AR, MALT1, NDRG1, and β-actin using immunoblot assays (top), as above. The quantitative data were expressed as the intensity of protein bands of the target proteins/β-actin relative to the control vehicle-treated group (bottom). (C) The mRNA levels of ARFL and ARv7 in the prostate cell lines, as indicated, were determined by RT-qPCR analysis (PZ: PZ-HPV-7, CA: CA-HPV-10). The mRNA levels of ARFL and ARv7 in 22Rv1 cells are regarded as 1, and ND represents non-detectable. * p < 0.05, ** p < 0.01.

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR

Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 5. Ectopic overexpression of ARv7 upregulates MALT1 expression to enhance NF-κB activation in PC-3 and 22Rv1 cells. An immunoblot assay of the expression of ARv7, MALT1, PSA, NDRG1, and β-actin in 22Rv1 (A) and PC-3 (B) cells. The quantitative data of the immunoblot assays were presented as the mean fold induction (±SE, n = 3) of the target proteins/β-actin relative to the mock- transfected group. The reporter activity of the MALT1 reporter vector was cotransfected with various concentrations of the ARv7 expression vectors, as indicated, for 24 h in the 22Rv1 (C) and PC-3 (E) cells. The data are presented as mean percentages (±SE, n = 6) relative to the control vehicle-treated group. The expression of ARv7, MALT1, IκBα, phospho-IκBα, p65, p50, Lamin B1, and GAPDH in the 22Rv1 (D) and PC-3 (F) cells after the separation of the nuclear and cytoplasmic fractions, as indicated, were determined by immunoblot assays. The numbers indicate the ratio of target proteins/GAPDH or Lamin B1 in relation to the mock-transfected group. The NF-κB (p65)-binding activity in the 22Rv1 (G) and PC-3 (H) cells after stably transfecting with ARv7. ** p < 0.01.

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Over Expression, Expressing, Activation Assay, Western Blot, Transfection, Activity Assay, Plasmid Preparation, Control, Binding Assay, Stable Transfection

Journal: International journal of molecular sciences

Article Title: Androgen Receptor Upregulates Mucosa-Associated Lymphoid Tissue 1 to Induce NF-κB Activity via Androgen-Dependent and -Independent Pathways in Prostate Carcinoma Cells.

doi: 10.3390/ijms24076245

Figure Lengend Snippet: Figure 7. Ectopic overexpression of ARv7 enhances tumorigenesis of PC-3 cells in xenograft mice models. The athymic male nude mice were subcutaneously injected with mock-overexpressed (PC-DNA) or ectopic-overexpressed ARv7 (PC-ARv7) cells for 25 days. (A) The average body weight (mean ± SE) of the mice during the experimental period. Photograph of the representative xenografted mice and tumors (B, top) and tumor sizes (B, bottom), which were measured every 3 days. The mice were sacrificed on the 25th day and the tumors were excised. (C) The quantitative data (mean ± SE) of the tumor weight of the PC-DNA and PC-ARv7 groups. (D) Whole-cell lysates of tumor samples from the PC-DNA and PC-ARv7 groups were subjected to immunoblot assays for ARv7, MALT1, NDRG1, and β-actin. (E) The quantitative data of the immunoblot assays are presented as the mean fold induction of ARv7, MALT1, and NDRG1 relative to the PC-DNA group. (F) The mRNA levels of ARv7, MALT1, and NDRG1 in the xenografted tumors were analyzed by RT-qPCR assays (±SE, n = 3). * p < 0.05, ** p < 0.01.

Article Snippet: The MALT1 shRNA lentiviral transduction particles (sc-35845-V, Santa Cruz Biotechnology, Santa Cruz, CA, USA) were transduced into LNCaP cells after the culture media was replaced with an RPMI-1640 medium plus 10% FBS and 5 μg/mL polybrene.

Techniques: Over Expression, Injection, Western Blot, Quantitative RT-PCR