Journal: Frontiers in Oncology

Article Title: HLA-BAT1 alters migration, invasion and pro-inflammatory cytokines in prostate cancer

doi: 10.3389/fonc.2022.969396

Figure Lengend Snippet: BAT1 Expression was decreased after siRNA Transfection and increased after cDNA Transfection in PC3 and 22RV1 cells. The samples were collected for a period of 24 hours. (A) Representative images and quantification of BAT1 protein expression in PC3 prostate cancer cells using western blot analysis showed a significant decrease in siBAT1 transfected cells when compared to control (*P<0.05). (B) Quantification of BAT1 RNA expression using RT-PCR in PC3 prostate cancer cells showed a significant decrease in BAT1 expression in siBAT1 transfected cells when compared to control (****P < 0.0001). (C) Representative images and quantification of BAT1 protein expression in PC3 prostate cancer cells using western blot analysis showed a significant increase in protein expression in BAT1cDNA when compared to control (***P < 0.001). (D) Quantification of BAT1 RNA expression using RT-PCR in PC3 prostate cancer cells showed a significant increase in BAT1 protein expression in BAT1cDNA cells when compared to control (**P < 0.01). (E) Representative images and quantification of BAT1 protein expression in 22RV1 prostate cancer cells using western blot analysis showed a significant decrease in siBAT1 transfected cells when compared to control (**P < 0.01). (F) Quantification of BAT1 RNA expression using RT-PCR in 22RV1 prostate cancer cells showed a significant decrease in BAT1 expression in siBAT1 transfected cells when compared to control (**P < 0.01) (G) Representative images and quantification of BAT1 protein expression in 22RV1 prostate cancer cells using western blot analysis showed a significant increase in BAT1cDNA when compared to control (***P < 0.001). (H) Quantification of BAT1 RNA expression using RT-PCR in 22RV1 prostate cancer cells showed a significant increase in BAT1 protein expression in BAT1cDNA cells when compared to control (**P < 0.01).

Article Snippet: Prostate cancer cells, PC3 and 22RV1, were seeded and transfected with BAT cDNA-GFP tagged lentiviral particles (BAT1cDNA) (pLenti-C-mGFP vector) (Origene, Rockville, MD, USA) using Polybrene transfection reagent (EMD Millipore, Burlington, MA) and OPTI-MEM Reduced Serum Medium.

Techniques: Expressing, Transfection, Western Blot, RNA Expression, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Oncology

Article Title: HLA-BAT1 alters migration, invasion and pro-inflammatory cytokines in prostate cancer

doi: 10.3389/fonc.2022.969396

Figure Lengend Snippet: BAT1 down-regulation increased PC3 prostate cancer cell migration and BAT1 overexpression decreased PC3 prostate cancer cell migration. (A) Representative images of Control PC3 and siBAT1 PC3 cells using 4X magnification. (B) Relative invasion of siBAT1 PC3 cells caused a significant increase in migration at 12hrs (**P < 0.01) and 24hrs (*P < 0.05) when compared to control. (C) Representative images of Control PC3 and BAT1cDNA PC3 cells using 4X magnification. (D) Relative invasion of (+) BAT1cDNA PC3 cells caused a significant decrease in migration at 12hrs (**P < 0.01) and 24hrs (***P < 0.001) when compared to control.

Article Snippet: Prostate cancer cells, PC3 and 22RV1, were seeded and transfected with BAT cDNA-GFP tagged lentiviral particles (BAT1cDNA) (pLenti-C-mGFP vector) (Origene, Rockville, MD, USA) using Polybrene transfection reagent (EMD Millipore, Burlington, MA) and OPTI-MEM Reduced Serum Medium.

Techniques: Migration, Over Expression

Journal: Frontiers in Oncology

Article Title: HLA-BAT1 alters migration, invasion and pro-inflammatory cytokines in prostate cancer

doi: 10.3389/fonc.2022.969396

Figure Lengend Snippet: BAT1 down-regulation increased PC3 and 22RV1 prostate cancer cell and BAT1 overexpression decreased PC3 and 22RV1 prostate cancer cell invasion. (A, B) Representative images of invasive siBAT1 PC3 and 22RV1 cells at a 4X magnification. (C) Relative invasion of siBAT1 PC3 cells caused a significant increase in invasion when compared to control at 12hrs (**P < 0.01) and 24hrs (****P < 0.0001). (D) Relative invasion of siBAT1 22RV1 cells caused a significant increase in invasion when compared to control at 12hrs (*P < 0.05) and 24hrs (***P < 0.001). (E, F) Representative images of invasive BAT1cDNA PC3 and 22RV1 cells at a 4X magnification. (G) Relative invasion of BAT1cDNA PC3 cells caused a significant decrease in invasion when compared to control at 12hrs (****P < 0.0001) and 24hrs (****P < 0.0001). (H) Relative invasion of BAT1cDNA 22RV1 cells caused a significant decrease in invasion when compared to control at 12hrs (***P < 0.001) and 24hrs (***P < 0.001).

Article Snippet: Prostate cancer cells, PC3 and 22RV1, were seeded and transfected with BAT cDNA-GFP tagged lentiviral particles (BAT1cDNA) (pLenti-C-mGFP vector) (Origene, Rockville, MD, USA) using Polybrene transfection reagent (EMD Millipore, Burlington, MA) and OPTI-MEM Reduced Serum Medium.

Techniques: Over Expression

Journal: Frontiers in Oncology

Article Title: HLA-BAT1 alters migration, invasion and pro-inflammatory cytokines in prostate cancer

doi: 10.3389/fonc.2022.969396

Figure Lengend Snippet: BAT1 expression showed changes in genes associated with inflammation, adhesion, and metastasis in PC3 and 22RV1 cells using qRT-PCR. The samples were collected for a period of 24 hours. (A, C) BAT1cDNA PC3 and 22RV1 cells showed a significant decrease in TNF-α expression when compared to siBAT1 (****P < 0.0001). (B, D) BAT1cDNA PC3 and 22RV1 cells showed a significant decrease in IL-6 expression when compared to siBAT1 (**P < 0.01) (***P < 0.001). (E) BAT1cDNA PC3 cells showed a significant decrease in MMP13 expression when compared to siBAT1 (****P < 0.0001). (F) BAT1cDNA 22RV1 cells showed a significant decrease in MMP10 expression when compared to siBAT1 (**P < 0.01). (G) BAT1cDNA 22RV1 cells showed a significant increase in TIMP2 expression when compared to siBAT1 (***P < 0.001).

Article Snippet: Prostate cancer cells, PC3 and 22RV1, were seeded and transfected with BAT cDNA-GFP tagged lentiviral particles (BAT1cDNA) (pLenti-C-mGFP vector) (Origene, Rockville, MD, USA) using Polybrene transfection reagent (EMD Millipore, Burlington, MA) and OPTI-MEM Reduced Serum Medium.

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Oncology

Article Title: HLA-BAT1 alters migration, invasion and pro-inflammatory cytokines in prostate cancer

doi: 10.3389/fonc.2022.969396

Figure Lengend Snippet: In vivo expression of BAT1, Ki67, TNF-α, IL-6, and MMP10. All of the samples were collected for a period of 24 hours. (A) Representative images of immunohistochemical staining BAT1 expression in mice prostate tumors previously injected with 22RV1 cells, BAT1cDNA 22RV1 cells, or shBAT1 22RV1 cells at a 20X (50μm) or 60X magnification (10μm) magnification. (B) Quantification mice prostate tumors obtained from BAT1cDNA mice prostate tumors showed a significant increase in BAT1 expression using immunohistochemistry when compared to control (*P < 0.05) and shBAT1 (**P < 0.01). (C) shBAT1 (**P < 0.01) and BAT1cDNA (*P < 0.05) tumors showed a significant decrease in Ki67 expression when compared to control. (D) shBAT1 tumors showed a significant increase in TNF-α expression when compared to control and BAT1cDNA (*P < 0.05). (E) shBAT1 tumors showed a significant increase in IL-6 expression when compared to control and BAT1cDNA mice prostate tumors (*P < 0.05). (F) shBAT1 tumors showed a significant increase in MMP10 expression when compared to control (*P < 0.05).

Article Snippet: Prostate cancer cells, PC3 and 22RV1, were seeded and transfected with BAT cDNA-GFP tagged lentiviral particles (BAT1cDNA) (pLenti-C-mGFP vector) (Origene, Rockville, MD, USA) using Polybrene transfection reagent (EMD Millipore, Burlington, MA) and OPTI-MEM Reduced Serum Medium.

Techniques: In Vivo, Expressing, Immunohistochemical staining, Staining, Injection, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Polycystin-1 Enhances Stemmness Potential of Umbilical Cord Blood-Derived Mesenchymal Stem Cells

doi: 10.3390/ijms22094868

Figure Lengend Snippet: Validation of PKD1 expression in transcript and protein level by PKD1 -expressing GFP/lentiviral vector. ( A ) GFP expression in UCB-MSC transfected with PKD1 -expressing vector. PKD1 expression ( B ) and quantitative data ( C ) in transcript level of PKD1 expressing UCB-MSC; ( D ) The expression of PKD1 at the protein level. Data shown represent the mean ± SD ( n = 3). ** p < 0.02 or *** p < 0.01 versus UCB-MSC only group.

Article Snippet: Each cell line was transfected with PKD1 overexpressing plasmid in the lentiviral GFP vector (Origene, Rockville, MD, USA) using TransIT-X2 (Mirus Bio, Madison, WI, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Plasmid Preparation, Transfection

Journal: Nature Communications

Article Title: Cooperative interaction between ERα and the EMT-inducer ZEB1 reprograms breast cancer cells for bone metastasis

doi: 10.1038/s41467-022-29723-5

Figure Lengend Snippet: a Immunoblots of an ERα and ZEB1 co-immunoprecipitation experiment. IPs with extracts from MCF7-V-ZEB1 without or with DOX treatment (for 1 week) were performed with antibodies specific to the exogenously expressed ZEB1 (left) or to the endogenous ERα (right). A control immunoprecipitation was performed in parallel with IgG, blotted, and exposed simultaneously (results are representative of n = 3 independent experiments). b , c Venn diagrams showing overlap of AP2γ ( b ) or FOXA1( c ) binding sites with ZEB1 and ERBSs from the ChIP-seq data. d Aggregation plot of the binding sites of ZEB1, ERα, AP2γ, FOXA1, GATA3, and P300, and the open chromatin histone marks H3K27ac, H3K4me1, and H3K9me3. Except for ERα, ChIP-seq data were from published data sets: GSE21234 (TFAP2C), GSE25315 (FOXA1), GSE60270 (GATA3, P300, H3K27ac, H3K4me1, and H3K9me3). e Genome browser snapshots of ZEB1, ERα, AP2γ, and FOXA1 enhancers of indicated genes. Highlights show shared binding sites for indicated factors. f Luciferase reporter assays with ERE-Luc in HEK293T cells infected with lentiviral constructs for shRNAs targeting FOXA1 , TFAP2C , or both mRNAs; scrambled shRNA (shScr) was used as negative control (mean ± SEM, n = 3 biologically independent experiments). g Co-IPs with HEK293T cells co-transfected with ZEB1 and ERα expression vectors. A control IP was performed with an IgG antibody (results are representative of n = 3 independent experiments). h Cells infected with viruses for expression of shScr, shTFAP2C, or shFOXA1. ERα ChIP-qPCR values are represented as the fold of the shScr in −DOX (mean ± SEM, n = 4 biologically independent experiments). i – j ERα ChIP-qPCR of binding sites associated with the genes LGALS1 and RAP1GAP2 ( i ), and SIRT5 and CD276 ( j ) in MCF7-V-ZEB1 cells infected with shScr or shTFAP2C. k ChIP-qPCR of ERBS at the TFAP2C 5′-UTR (means ± SEM, n = 4 and n = 3 biologically independent experiments in i and k , and j , respectively). l Immunoblots of extracts from MCF7-V-ZEB1 cells. veh, E2, and FI stand for vehicle, 17β-estradiol, and forskolin + IBMX, respectively. For bar graphs, p values are indicated above the bars; statistical significance was determined with a two-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: The lentiviral UBC-GFP-T2A-Luciferase dual reporter for in vivo imaging and the pMDLg and pRSV-Rev packaging plasmids were from BioCat GmbH.

Techniques: Western Blot, Immunoprecipitation, Binding Assay, ChIP-sequencing, Luciferase, Infection, Construct, shRNA, Negative Control, Transfection, Expressing