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96
Integrated DNA Technologies sirna targeting sult2b1
(A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates SULT2B1b KD. (C) Number of <t>SULT2B1</t> reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.
Sirna Targeting Sult2b1, supplied by Integrated DNA Technologies, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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86
Genechem mouse sult2b1 small interfering rna
Primer sets used for qPCR.
Mouse Sult2b1 Small Interfering Rna, supplied by Genechem, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Genechem human sult2b1 small interfering rna
Primer sets used for qPCR.
Human Sult2b1 Small Interfering Rna, supplied by Genechem, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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86
Millipore sult2b1 mission shrna lentiviral particles
AR levels, invasion phenotype and mechanical changes of SULT2B-ablated CRPC cells. (A) SULT2B and AR levels in CRISPR’d clones and parent C4-2 cells. (B) <t>SULT2B1</t> mRNAs in CRISPR’d clones. Bar charts show average ± SEM of three biological replicates in duplicate qRT-PCR assay. **P < .001. (C). Matrigel invasion of KO and non-KO cells in Transwell chamber assay. Photomicrographs representative of two biological replicates, showing Matrigel-invaded cells, stained with crystal violet at 72 hours post culture. (D,E) AFM analysis of KO, non-KO and parent C4-2 cells. (D) Young’s modulus measuring cell stiffness (in units of Pascal, Pa); (E) AFM tip to cell bonding in units of Newton force N, which measures cell adhesion. Each point is a mode of parameter values measured over a single cell area. Three biological replicates per cell line were analyzed. Number of live cells analyzed for each cell line: 25 (parent C4-2); 31 (clone #21); 23 (clone #23); 30 (clone #3); 31 (clone #6). Horizontal lines are mean values and whiskers are standard deviations. (F,G). SULT2B (F) and AR (G) levels in SULT2B KD cells. GAPDH (a cytosolic protein) and Ku86 (a nuclear protein) are internal controls. Size markers verified molecular weights of immunoblotted proteins.
Sult2b1 Mission Shrna Lentiviral Particles, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Qiagen sult2b1 sirna
AR levels, invasion phenotype and mechanical changes of SULT2B-ablated CRPC cells. (A) SULT2B and AR levels in CRISPR’d clones and parent C4-2 cells. (B) <t>SULT2B1</t> mRNAs in CRISPR’d clones. Bar charts show average ± SEM of three biological replicates in duplicate qRT-PCR assay. **P < .001. (C). Matrigel invasion of KO and non-KO cells in Transwell chamber assay. Photomicrographs representative of two biological replicates, showing Matrigel-invaded cells, stained with crystal violet at 72 hours post culture. (D,E) AFM analysis of KO, non-KO and parent C4-2 cells. (D) Young’s modulus measuring cell stiffness (in units of Pascal, Pa); (E) AFM tip to cell bonding in units of Newton force N, which measures cell adhesion. Each point is a mode of parameter values measured over a single cell area. Three biological replicates per cell line were analyzed. Number of live cells analyzed for each cell line: 25 (parent C4-2); 31 (clone #21); 23 (clone #23); 30 (clone #3); 31 (clone #6). Horizontal lines are mean values and whiskers are standard deviations. (F,G). SULT2B (F) and AR (G) levels in SULT2B KD cells. GAPDH (a cytosolic protein) and Ku86 (a nuclear protein) are internal controls. Size markers verified molecular weights of immunoblotted proteins.
Sult2b1 Sirna, supplied by Qiagen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Ribobio co human sult2b1 specific sirna duplex oligo ribonucleotides
<t>SULT2B1-deficient</t> mice were susceptible to gastric tumors upon induction with the carcinogenic agent 3-MCA. a SULT2B1 expression in human normal gastric mucosal tissue was studied by immunohistochemical staining and Western blotting analysis. b The SULT2B1 mRNA level in different murine organs was detected by qRT-PCR. c The SULT2B1 mRNA expression level in the gastric tissues of WT and SULT2B1 −/− mice was measured by qRT-PCR. The concentration of cholesterol sulfate in the gastric tissues of WT and SULT2B1 −/− mice was detected by LC-MS. d The gastric carcinoma model was established by 3-MCA induction in WT and SULT2B1 −/− mice. e Three months after induction, the number of tumor-bearing mice with or without bloody ascites in the WT and SULT2B1 −/− mice was plotted. f The survival probabilities of WT and SULT2B1 −/− mice upon 3-MCA treatment. g The results from the H&E staining and immunohistochemical staining for CK8, CKpan, and VIM expression in the nodules are presented. * P < 0.05, ** P < 0.01
Human Sult2b1 Specific Sirna Duplex Oligo Ribonucleotides, supplied by Ribobio co, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/human sult2b1 specific sirna duplex oligo ribonucleotides/product/Ribobio co
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86
fluidigm sult2b1 sirna
(A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates <t>SULT2B1b</t> KD. (C) Number of <t>SULT2B1</t> reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.
Sult2b1 Sirna, supplied by fluidigm, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sult2b1 sirna/product/fluidigm
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Image Search Results


(A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates SULT2B1b KD. (C) Number of SULT2B1 reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: (A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates SULT2B1b KD. (C) Number of SULT2B1 reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.

Article Snippet: RNAi studies were completed using control/non-targeting siRNA complexes (Dharmacon) and siRNA targeting SULT2B1 (IDT, HSC.RNAI.N004605.12.2), TNF (IDT, hs.Ri.TNF.13.2), TL1 (IDT, hs.Ri.TNFSF15.13.3), or DAXX (IDT, hs.Ri.DAXX.13.1) using Lipofectamine RNAiMax (ThermoFisher), according the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Transfection

(A) Gene co-expression indicates SULT2B1 negatively correlates with TNF, CD40LG, TRADD, and TRAF2 in bone marrow-derived metastatic prostate cancer samples from individuals with no prior treatment. (B) Co-expression of SULT2B1 with NFKB1, FADD, and DR4 determines significant negative correlations with each of these genes in lymph node-derived CRPC samples. (C) Gene co-expression of SULT2B1 with RANKL demonstrates a strong positive correlation in bone marrow-derived prostate cancer samples. Pearson correlation coefficients (r) and p-values determined by a permutation test (p) are indicated for each correlation plot. Dots indicate samples obtained from individual patients.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: (A) Gene co-expression indicates SULT2B1 negatively correlates with TNF, CD40LG, TRADD, and TRAF2 in bone marrow-derived metastatic prostate cancer samples from individuals with no prior treatment. (B) Co-expression of SULT2B1 with NFKB1, FADD, and DR4 determines significant negative correlations with each of these genes in lymph node-derived CRPC samples. (C) Gene co-expression of SULT2B1 with RANKL demonstrates a strong positive correlation in bone marrow-derived prostate cancer samples. Pearson correlation coefficients (r) and p-values determined by a permutation test (p) are indicated for each correlation plot. Dots indicate samples obtained from individual patients.

Article Snippet: RNAi studies were completed using control/non-targeting siRNA complexes (Dharmacon) and siRNA targeting SULT2B1 (IDT, HSC.RNAI.N004605.12.2), TNF (IDT, hs.Ri.TNF.13.2), TL1 (IDT, hs.Ri.TNFSF15.13.3), or DAXX (IDT, hs.Ri.DAXX.13.1) using Lipofectamine RNAiMax (ThermoFisher), according the manufacturer’s instructions.

Techniques: Expressing, Derivative Assay

Primer sets used for qPCR.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: Primer sets used for qPCR.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques:

The mRNA (A) and protein (B) levels of SULT2B1 in transduced Hepa1-6 cells were detected by qPCR and western blot analysis respectively. SULT2B1 sulfotransferase activity in transduced Hepa1-6 cells was detected by sulfotransferase assay in vitro (C) or by measuring the conversion rate of [ 3 H] cholesterol to [ 3 H] methanol-water-soluble counts by adding [ 3 H] cholesterol to live cells (D). SULT2B1b protein expression in Hepa1-6 cells transduced with Ad-EGFP or Ad-SULT2B1b (E). CCK-8 assay of the growth curves of Hepa1-6 cells transduced with SULT2B1-RNAi-LV, Ad-SULT2B1b, or vector controls (F,G). *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA (A) and protein (B) levels of SULT2B1 in transduced Hepa1-6 cells were detected by qPCR and western blot analysis respectively. SULT2B1 sulfotransferase activity in transduced Hepa1-6 cells was detected by sulfotransferase assay in vitro (C) or by measuring the conversion rate of [ 3 H] cholesterol to [ 3 H] methanol-water-soluble counts by adding [ 3 H] cholesterol to live cells (D). SULT2B1b protein expression in Hepa1-6 cells transduced with Ad-EGFP or Ad-SULT2B1b (E). CCK-8 assay of the growth curves of Hepa1-6 cells transduced with SULT2B1-RNAi-LV, Ad-SULT2B1b, or vector controls (F,G). *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Western Blot, Activity Assay, In Vitro, Expressing, Transduction, CCK-8 Assay, Plasmid Preparation

(A) Flow cytometry analysis of the percentage of cells in different phases of the cell cycle in untransduced Hepa1-6 cells or Hepa1-6 cells transduced with vector control or SULT2B1-RNAi-LV analyzed with propidium iodide staining. (B) Flow cytometry analysis of the percentage of apoptotic cells in untreated Hepa1-6 cells or Hepa1-6 cells treated with vector control or SULT2B1-RNAi-LV as analyzed by Annexin V-APC and 7-ADD staining. The data represents the mean of three independent experiments ± S.D. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: (A) Flow cytometry analysis of the percentage of cells in different phases of the cell cycle in untransduced Hepa1-6 cells or Hepa1-6 cells transduced with vector control or SULT2B1-RNAi-LV analyzed with propidium iodide staining. (B) Flow cytometry analysis of the percentage of apoptotic cells in untreated Hepa1-6 cells or Hepa1-6 cells treated with vector control or SULT2B1-RNAi-LV as analyzed by Annexin V-APC and 7-ADD staining. The data represents the mean of three independent experiments ± S.D. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Flow Cytometry, Transduction, Plasmid Preparation, Staining

The mRNA levels of FAS and BCL2 following SULT2B1b overexpression (A, C) and SULT2B1b inhibition (B, D) were analyzed by qPCR. Western blot analysis of: (E) FAS and BCL2 protein levels in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV upon serum starvation at 6 h, 12 h and 24 h. (F) The protein levels of BCL2 and MYC in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV at different multiplicity of infections (MOI = 25, 50, 100) and cultured in serum-deprived medium for 24 h. (G) FAS and BCL2 protein levels in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells cultured in serum-free medium containing 10 ng/mL TNF-α and 10 µg/mL CHX for the indicated times. (H) The BCL2 and MYC protein levels in Hepa1-6 cells infected with NC-GFP-LV and SULT2B1-RNAi-LV at MOI of 25, 50, 100 and treated with 10 ng/mL TNF-α and 10 µg/mL CHX. *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA levels of FAS and BCL2 following SULT2B1b overexpression (A, C) and SULT2B1b inhibition (B, D) were analyzed by qPCR. Western blot analysis of: (E) FAS and BCL2 protein levels in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV upon serum starvation at 6 h, 12 h and 24 h. (F) The protein levels of BCL2 and MYC in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV at different multiplicity of infections (MOI = 25, 50, 100) and cultured in serum-deprived medium for 24 h. (G) FAS and BCL2 protein levels in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells cultured in serum-free medium containing 10 ng/mL TNF-α and 10 µg/mL CHX for the indicated times. (H) The BCL2 and MYC protein levels in Hepa1-6 cells infected with NC-GFP-LV and SULT2B1-RNAi-LV at MOI of 25, 50, 100 and treated with 10 ng/mL TNF-α and 10 µg/mL CHX. *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Over Expression, Inhibition, Western Blot, Cell Culture, Infection

The mRNA (A) and protein (B) levels of cyclinB1 were analyzed by qPCR and Western-blot, respectively. (C) The stability of cyclinB1 protein in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells was determined with 100 µg/ml CHX treatment. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA (A) and protein (B) levels of cyclinB1 were analyzed by qPCR and Western-blot, respectively. (C) The stability of cyclinB1 protein in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells was determined with 100 µg/ml CHX treatment. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Western Blot

1.5×10 6 NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells were injected subcutaneously into the subaxillary space of each nude mouse. (A) The growth curve of xenografts. (B) Representative fluorescence images of xenografts. (C) Images of dissected tumors. (D) The weights of dissected tumors. (E) Western-blot analysis of the protein levels of the cell growth-associated genes, BCL2, MYC, cyclinD1, and cyclinB1 in dissected tumors. The quantification of protein levels of the above genes is shown to the right. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: 1.5×10 6 NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells were injected subcutaneously into the subaxillary space of each nude mouse. (A) The growth curve of xenografts. (B) Representative fluorescence images of xenografts. (C) Images of dissected tumors. (D) The weights of dissected tumors. (E) Western-blot analysis of the protein levels of the cell growth-associated genes, BCL2, MYC, cyclinD1, and cyclinB1 in dissected tumors. The quantification of protein levels of the above genes is shown to the right. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Injection, Fluorescence, Western Blot

Endogenous expression of SULT2B1b mRNA (A) and protein levels (B) was measured by qPCR and Western blot assay, respectively. (C) Proliferation growth curves of the human hepatocarcinoma cell lines as measured by the CCK-8 assay. (D) Pearson correlation and simple linear regression analysis of SULT2B1 mRNA levels with cell proliferation (r = 0.931, R 2 = 0.867, y = 0.9386×−0.2832). (E) Expressions of SULT2B1 mRNA levels in para-tumor and tumor tissues of clinical human hepatocarcinoma samples was detected by qPCR (n = 6). The mRNA levels were normalized to the internal control and represented as the means of results from different samples ± standard error (SE). The PCR products were visualized on a 2% agarose gel containing 5 mg/ml ethidium bromide,GADPH was used as internal control. *represents P <0.05 vs. para-tumor tissues.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: Endogenous expression of SULT2B1b mRNA (A) and protein levels (B) was measured by qPCR and Western blot assay, respectively. (C) Proliferation growth curves of the human hepatocarcinoma cell lines as measured by the CCK-8 assay. (D) Pearson correlation and simple linear regression analysis of SULT2B1 mRNA levels with cell proliferation (r = 0.931, R 2 = 0.867, y = 0.9386×−0.2832). (E) Expressions of SULT2B1 mRNA levels in para-tumor and tumor tissues of clinical human hepatocarcinoma samples was detected by qPCR (n = 6). The mRNA levels were normalized to the internal control and represented as the means of results from different samples ± standard error (SE). The PCR products were visualized on a 2% agarose gel containing 5 mg/ml ethidium bromide,GADPH was used as internal control. *represents P <0.05 vs. para-tumor tissues.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Expressing, Western Blot, CCK-8 Assay, Agarose Gel Electrophoresis

(A) qPCR analysis of SULT2B1b mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1 or SULT2B1b-specific siRNA treatment. Cell proliferation rates of BEL-7402 (B) and SMMC-7721 (C) cells treated with SULT2B1 or SULT2B1b-specific siRNA was assessed by CCK-8 assay. (D) cyclinB1 mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment was detected by qPCR assay. (E) cyclinB1 protein levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment detected by Western-blot analysis, β-actin as internal control. * P <0.05 vs. NC group . BEL-7402 cells (1×10 6 ) infected with NC-RFP-LV or hSULT2B1-RNAi-LV were injected subcutaneously into right subaxillary region of each nude mouse. The xenograft tumor growth curve (F), representative images (G) and the weights (H) of dissected xenografts were shown. * P <0.05 vs. NC-RFP-LV control group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: (A) qPCR analysis of SULT2B1b mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1 or SULT2B1b-specific siRNA treatment. Cell proliferation rates of BEL-7402 (B) and SMMC-7721 (C) cells treated with SULT2B1 or SULT2B1b-specific siRNA was assessed by CCK-8 assay. (D) cyclinB1 mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment was detected by qPCR assay. (E) cyclinB1 protein levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment detected by Western-blot analysis, β-actin as internal control. * P <0.05 vs. NC group . BEL-7402 cells (1×10 6 ) infected with NC-RFP-LV or hSULT2B1-RNAi-LV were injected subcutaneously into right subaxillary region of each nude mouse. The xenograft tumor growth curve (F), representative images (G) and the weights (H) of dissected xenografts were shown. * P <0.05 vs. NC-RFP-LV control group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: CCK-8 Assay, Western Blot, Infection, Injection

Primer sets used for qPCR.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: Primer sets used for qPCR.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques:

The mRNA (A) and protein (B) levels of SULT2B1 in transduced Hepa1-6 cells were detected by qPCR and western blot analysis respectively. SULT2B1 sulfotransferase activity in transduced Hepa1-6 cells was detected by sulfotransferase assay in vitro (C) or by measuring the conversion rate of [ 3 H] cholesterol to [ 3 H] methanol-water-soluble counts by adding [ 3 H] cholesterol to live cells (D). SULT2B1b protein expression in Hepa1-6 cells transduced with Ad-EGFP or Ad-SULT2B1b (E). CCK-8 assay of the growth curves of Hepa1-6 cells transduced with SULT2B1-RNAi-LV, Ad-SULT2B1b, or vector controls (F,G). *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA (A) and protein (B) levels of SULT2B1 in transduced Hepa1-6 cells were detected by qPCR and western blot analysis respectively. SULT2B1 sulfotransferase activity in transduced Hepa1-6 cells was detected by sulfotransferase assay in vitro (C) or by measuring the conversion rate of [ 3 H] cholesterol to [ 3 H] methanol-water-soluble counts by adding [ 3 H] cholesterol to live cells (D). SULT2B1b protein expression in Hepa1-6 cells transduced with Ad-EGFP or Ad-SULT2B1b (E). CCK-8 assay of the growth curves of Hepa1-6 cells transduced with SULT2B1-RNAi-LV, Ad-SULT2B1b, or vector controls (F,G). *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Western Blot, Activity Assay, In Vitro, Expressing, Transduction, CCK-8 Assay, Plasmid Preparation

(A) Flow cytometry analysis of the percentage of cells in different phases of the cell cycle in untransduced Hepa1-6 cells or Hepa1-6 cells transduced with vector control or SULT2B1-RNAi-LV analyzed with propidium iodide staining. (B) Flow cytometry analysis of the percentage of apoptotic cells in untreated Hepa1-6 cells or Hepa1-6 cells treated with vector control or SULT2B1-RNAi-LV as analyzed by Annexin V-APC and 7-ADD staining. The data represents the mean of three independent experiments ± S.D. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: (A) Flow cytometry analysis of the percentage of cells in different phases of the cell cycle in untransduced Hepa1-6 cells or Hepa1-6 cells transduced with vector control or SULT2B1-RNAi-LV analyzed with propidium iodide staining. (B) Flow cytometry analysis of the percentage of apoptotic cells in untreated Hepa1-6 cells or Hepa1-6 cells treated with vector control or SULT2B1-RNAi-LV as analyzed by Annexin V-APC and 7-ADD staining. The data represents the mean of three independent experiments ± S.D. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Flow Cytometry, Transduction, Plasmid Preparation, Staining

The mRNA levels of FAS and BCL2 following SULT2B1b overexpression (A, C) and SULT2B1b inhibition (B, D) were analyzed by qPCR. Western blot analysis of: (E) FAS and BCL2 protein levels in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV upon serum starvation at 6 h, 12 h and 24 h. (F) The protein levels of BCL2 and MYC in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV at different multiplicity of infections (MOI = 25, 50, 100) and cultured in serum-deprived medium for 24 h. (G) FAS and BCL2 protein levels in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells cultured in serum-free medium containing 10 ng/mL TNF-α and 10 µg/mL CHX for the indicated times. (H) The BCL2 and MYC protein levels in Hepa1-6 cells infected with NC-GFP-LV and SULT2B1-RNAi-LV at MOI of 25, 50, 100 and treated with 10 ng/mL TNF-α and 10 µg/mL CHX. *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA levels of FAS and BCL2 following SULT2B1b overexpression (A, C) and SULT2B1b inhibition (B, D) were analyzed by qPCR. Western blot analysis of: (E) FAS and BCL2 protein levels in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV upon serum starvation at 6 h, 12 h and 24 h. (F) The protein levels of BCL2 and MYC in Hepa1-6 cells treated with NC-GFP-LV or SULT2B1-RNAi-LV at different multiplicity of infections (MOI = 25, 50, 100) and cultured in serum-deprived medium for 24 h. (G) FAS and BCL2 protein levels in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells cultured in serum-free medium containing 10 ng/mL TNF-α and 10 µg/mL CHX for the indicated times. (H) The BCL2 and MYC protein levels in Hepa1-6 cells infected with NC-GFP-LV and SULT2B1-RNAi-LV at MOI of 25, 50, 100 and treated with 10 ng/mL TNF-α and 10 µg/mL CHX. *represents P <0.05 vs. Ad-GFP group or NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Over Expression, Inhibition, Western Blot, Cell Culture, Infection

The mRNA (A) and protein (B) levels of cyclinB1 were analyzed by qPCR and Western-blot, respectively. (C) The stability of cyclinB1 protein in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells was determined with 100 µg/ml CHX treatment. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: The mRNA (A) and protein (B) levels of cyclinB1 were analyzed by qPCR and Western-blot, respectively. (C) The stability of cyclinB1 protein in NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells was determined with 100 µg/ml CHX treatment. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Western Blot

1.5×10 6 NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells were injected subcutaneously into the subaxillary space of each nude mouse. (A) The growth curve of xenografts. (B) Representative fluorescence images of xenografts. (C) Images of dissected tumors. (D) The weights of dissected tumors. (E) Western-blot analysis of the protein levels of the cell growth-associated genes, BCL2, MYC, cyclinD1, and cyclinB1 in dissected tumors. The quantification of protein levels of the above genes is shown to the right. *represents P <0.05 vs. NC-GFP-LV group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: 1.5×10 6 NC-GFP-LV or SULT2B1-RNAi-LV Hepa1-6 cells were injected subcutaneously into the subaxillary space of each nude mouse. (A) The growth curve of xenografts. (B) Representative fluorescence images of xenografts. (C) Images of dissected tumors. (D) The weights of dissected tumors. (E) Western-blot analysis of the protein levels of the cell growth-associated genes, BCL2, MYC, cyclinD1, and cyclinB1 in dissected tumors. The quantification of protein levels of the above genes is shown to the right. *represents P <0.05 vs. NC-GFP-LV group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Injection, Fluorescence, Western Blot

Endogenous expression of SULT2B1b mRNA (A) and protein levels (B) was measured by qPCR and Western blot assay, respectively. (C) Proliferation growth curves of the human hepatocarcinoma cell lines as measured by the CCK-8 assay. (D) Pearson correlation and simple linear regression analysis of SULT2B1 mRNA levels with cell proliferation (r = 0.931, R 2 = 0.867, y = 0.9386×−0.2832). (E) Expressions of SULT2B1 mRNA levels in para-tumor and tumor tissues of clinical human hepatocarcinoma samples was detected by qPCR (n = 6). The mRNA levels were normalized to the internal control and represented as the means of results from different samples ± standard error (SE). The PCR products were visualized on a 2% agarose gel containing 5 mg/ml ethidium bromide,GADPH was used as internal control. *represents P <0.05 vs. para-tumor tissues.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: Endogenous expression of SULT2B1b mRNA (A) and protein levels (B) was measured by qPCR and Western blot assay, respectively. (C) Proliferation growth curves of the human hepatocarcinoma cell lines as measured by the CCK-8 assay. (D) Pearson correlation and simple linear regression analysis of SULT2B1 mRNA levels with cell proliferation (r = 0.931, R 2 = 0.867, y = 0.9386×−0.2832). (E) Expressions of SULT2B1 mRNA levels in para-tumor and tumor tissues of clinical human hepatocarcinoma samples was detected by qPCR (n = 6). The mRNA levels were normalized to the internal control and represented as the means of results from different samples ± standard error (SE). The PCR products were visualized on a 2% agarose gel containing 5 mg/ml ethidium bromide,GADPH was used as internal control. *represents P <0.05 vs. para-tumor tissues.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: Expressing, Western Blot, CCK-8 Assay, Agarose Gel Electrophoresis

(A) qPCR analysis of SULT2B1b mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1 or SULT2B1b-specific siRNA treatment. Cell proliferation rates of BEL-7402 (B) and SMMC-7721 (C) cells treated with SULT2B1 or SULT2B1b-specific siRNA was assessed by CCK-8 assay. (D) cyclinB1 mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment was detected by qPCR assay. (E) cyclinB1 protein levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment detected by Western-blot analysis, β-actin as internal control. * P <0.05 vs. NC group . BEL-7402 cells (1×10 6 ) infected with NC-RFP-LV or hSULT2B1-RNAi-LV were injected subcutaneously into right subaxillary region of each nude mouse. The xenograft tumor growth curve (F), representative images (G) and the weights (H) of dissected xenografts were shown. * P <0.05 vs. NC-RFP-LV control group.

Journal: PLoS ONE

Article Title: Hydroxysteroid Sulfotransferase SULT2B1b Promotes Hepatocellular Carcinoma Cells Proliferation In Vitro and In Vivo

doi: 10.1371/journal.pone.0060853

Figure Lengend Snippet: (A) qPCR analysis of SULT2B1b mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1 or SULT2B1b-specific siRNA treatment. Cell proliferation rates of BEL-7402 (B) and SMMC-7721 (C) cells treated with SULT2B1 or SULT2B1b-specific siRNA was assessed by CCK-8 assay. (D) cyclinB1 mRNA levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment was detected by qPCR assay. (E) cyclinB1 protein levels in BEL-7402 and SMMC-7721 cells with SULT2B1b-specific siRNA treatment detected by Western-blot analysis, β-actin as internal control. * P <0.05 vs. NC group . BEL-7402 cells (1×10 6 ) infected with NC-RFP-LV or hSULT2B1-RNAi-LV were injected subcutaneously into right subaxillary region of each nude mouse. The xenograft tumor growth curve (F), representative images (G) and the weights (H) of dissected xenografts were shown. * P <0.05 vs. NC-RFP-LV control group.

Article Snippet: Mouse SULT2B1 small interfering RNA (mSULT2B1-RNAi-LV, target sequence, CAGTGTTTACCGAGAGCAAAT ; TU = 1.5×10 9 /mL), human SULT2B1 small interfering RNA (hSULT2B1-RNAi-LV, target sequence: GGGACTTCCTCAAAGGCGA ; TU = 3×10 8 /mL) were prepared by Genechem corporation (Shanghai, China).

Techniques: CCK-8 Assay, Western Blot, Infection, Injection

AR levels, invasion phenotype and mechanical changes of SULT2B-ablated CRPC cells. (A) SULT2B and AR levels in CRISPR’d clones and parent C4-2 cells. (B) SULT2B1 mRNAs in CRISPR’d clones. Bar charts show average ± SEM of three biological replicates in duplicate qRT-PCR assay. **P < .001. (C). Matrigel invasion of KO and non-KO cells in Transwell chamber assay. Photomicrographs representative of two biological replicates, showing Matrigel-invaded cells, stained with crystal violet at 72 hours post culture. (D,E) AFM analysis of KO, non-KO and parent C4-2 cells. (D) Young’s modulus measuring cell stiffness (in units of Pascal, Pa); (E) AFM tip to cell bonding in units of Newton force N, which measures cell adhesion. Each point is a mode of parameter values measured over a single cell area. Three biological replicates per cell line were analyzed. Number of live cells analyzed for each cell line: 25 (parent C4-2); 31 (clone #21); 23 (clone #23); 30 (clone #3); 31 (clone #6). Horizontal lines are mean values and whiskers are standard deviations. (F,G). SULT2B (F) and AR (G) levels in SULT2B KD cells. GAPDH (a cytosolic protein) and Ku86 (a nuclear protein) are internal controls. Size markers verified molecular weights of immunoblotted proteins.

Journal: Endocrinology

Article Title: Inhibitory Interplay of SULT2B1b Sulfotransferase with AKR1C3 Aldo-keto Reductase in Prostate Cancer

doi: 10.1210/endocr/bqz042

Figure Lengend Snippet: AR levels, invasion phenotype and mechanical changes of SULT2B-ablated CRPC cells. (A) SULT2B and AR levels in CRISPR’d clones and parent C4-2 cells. (B) SULT2B1 mRNAs in CRISPR’d clones. Bar charts show average ± SEM of three biological replicates in duplicate qRT-PCR assay. **P < .001. (C). Matrigel invasion of KO and non-KO cells in Transwell chamber assay. Photomicrographs representative of two biological replicates, showing Matrigel-invaded cells, stained with crystal violet at 72 hours post culture. (D,E) AFM analysis of KO, non-KO and parent C4-2 cells. (D) Young’s modulus measuring cell stiffness (in units of Pascal, Pa); (E) AFM tip to cell bonding in units of Newton force N, which measures cell adhesion. Each point is a mode of parameter values measured over a single cell area. Three biological replicates per cell line were analyzed. Number of live cells analyzed for each cell line: 25 (parent C4-2); 31 (clone #21); 23 (clone #23); 30 (clone #3); 31 (clone #6). Horizontal lines are mean values and whiskers are standard deviations. (F,G). SULT2B (F) and AR (G) levels in SULT2B KD cells. GAPDH (a cytosolic protein) and Ku86 (a nuclear protein) are internal controls. Size markers verified molecular weights of immunoblotted proteins.

Article Snippet: For stable SULT2B KD, C4-2B cells were infected with SULT2B1 MISSION® shRNA lentiviral particles (SHCLNV- {"type":"entrez-nucleotide","attrs":{"text":"NM_004605","term_id":"31563387","term_text":"NM_004605"}} NM_004605 , Sigma-Aldrich) and puromycin-resistant clonal lines were isolated.

Techniques: Clone Assay, Quantitative RT-PCR, Transwell Chamber Assay, Staining

SULT2B1-deficient mice were susceptible to gastric tumors upon induction with the carcinogenic agent 3-MCA. a SULT2B1 expression in human normal gastric mucosal tissue was studied by immunohistochemical staining and Western blotting analysis. b The SULT2B1 mRNA level in different murine organs was detected by qRT-PCR. c The SULT2B1 mRNA expression level in the gastric tissues of WT and SULT2B1 −/− mice was measured by qRT-PCR. The concentration of cholesterol sulfate in the gastric tissues of WT and SULT2B1 −/− mice was detected by LC-MS. d The gastric carcinoma model was established by 3-MCA induction in WT and SULT2B1 −/− mice. e Three months after induction, the number of tumor-bearing mice with or without bloody ascites in the WT and SULT2B1 −/− mice was plotted. f The survival probabilities of WT and SULT2B1 −/− mice upon 3-MCA treatment. g The results from the H&E staining and immunohistochemical staining for CK8, CKpan, and VIM expression in the nodules are presented. * P < 0.05, ** P < 0.01

Journal: Lipids in Health and Disease

Article Title: Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent

doi: 10.1186/s12944-019-1149-6

Figure Lengend Snippet: SULT2B1-deficient mice were susceptible to gastric tumors upon induction with the carcinogenic agent 3-MCA. a SULT2B1 expression in human normal gastric mucosal tissue was studied by immunohistochemical staining and Western blotting analysis. b The SULT2B1 mRNA level in different murine organs was detected by qRT-PCR. c The SULT2B1 mRNA expression level in the gastric tissues of WT and SULT2B1 −/− mice was measured by qRT-PCR. The concentration of cholesterol sulfate in the gastric tissues of WT and SULT2B1 −/− mice was detected by LC-MS. d The gastric carcinoma model was established by 3-MCA induction in WT and SULT2B1 −/− mice. e Three months after induction, the number of tumor-bearing mice with or without bloody ascites in the WT and SULT2B1 −/− mice was plotted. f The survival probabilities of WT and SULT2B1 −/− mice upon 3-MCA treatment. g The results from the H&E staining and immunohistochemical staining for CK8, CKpan, and VIM expression in the nodules are presented. * P < 0.05, ** P < 0.01

Article Snippet: The cells were transfected with human SULT2B1-specific siRNA duplex oligo ribonucleotides, with a 5′-GATCGAGATCATCTGCTTA-3′ targeting sequence, or negative control duplexes (RiboBio, Guangzhou, China) using Lipofectamine 3000.

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Quantitative RT-PCR, Concentration Assay, Liquid Chromatography with Mass Spectroscopy

RNA-seq analysis indicated that SULT2B1 is involved in epithelial development. a Schematic diagram of the SULT2B1 gene (blue bars, exons) and the CRISPR/CAS9 gRNA sequence (guide a and b , red). TA cloning results of clone 7 showed that both alleles of the human SULT2B1 gene were modified on exon 2 by the CRISPR/CAS9 system. b The SULT2B1 −/− GES-1 cell line was established by the CRISPR/CAS9 technique. SULT2B1 protein expression was measured by Western blotting analysis in GES-1 cells, SULT2B1 −/− GES-1 cells, and GES-1 cells transfected for 48 h with Ad-GFP or Ad-SULT2B1b. c Among the genes changed by both SULT2B1b overexpression and SULT2B1 deletion, some genes in epithelial development were enriched. The fold changes in these gene levels are shown by the heatmap

Journal: Lipids in Health and Disease

Article Title: Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent

doi: 10.1186/s12944-019-1149-6

Figure Lengend Snippet: RNA-seq analysis indicated that SULT2B1 is involved in epithelial development. a Schematic diagram of the SULT2B1 gene (blue bars, exons) and the CRISPR/CAS9 gRNA sequence (guide a and b , red). TA cloning results of clone 7 showed that both alleles of the human SULT2B1 gene were modified on exon 2 by the CRISPR/CAS9 system. b The SULT2B1 −/− GES-1 cell line was established by the CRISPR/CAS9 technique. SULT2B1 protein expression was measured by Western blotting analysis in GES-1 cells, SULT2B1 −/− GES-1 cells, and GES-1 cells transfected for 48 h with Ad-GFP or Ad-SULT2B1b. c Among the genes changed by both SULT2B1b overexpression and SULT2B1 deletion, some genes in epithelial development were enriched. The fold changes in these gene levels are shown by the heatmap

Article Snippet: The cells were transfected with human SULT2B1-specific siRNA duplex oligo ribonucleotides, with a 5′-GATCGAGATCATCTGCTTA-3′ targeting sequence, or negative control duplexes (RiboBio, Guangzhou, China) using Lipofectamine 3000.

Techniques: RNA Sequencing Assay, CRISPR, Sequencing, TA Cloning, Modification, Expressing, Western Blot, Transfection, Over Expression

SULT2B1 deletion suppressed PI3K/AKT signaling in GES-1 cells and contributed to transformation upon 3-MCA induction. a , b The GES-1 and SULT2B1 −/− GES-1 cells were stimulated with IGF-1 (100 ng/mL, a ) or EGF (100 ng/mL, b ) for 5, 15, 30, 60, or 180 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. c GES-1 and SULT2B1 −/− GES-1 cells were stimulated with IGF-1 (100 ng/mL) or EGF (100 ng/mL) for 8 h. The mRNA levels of the genes involved in gastric epithelial function were measured by qRT-PCR. d Immunofluorescent staining of claudin-1 in GES-1 and SULT2B1 −/− GES-1 cells. The nuclei were revealed by DAPI staining (blue). e Wound healing assay of GES-1, SULT2B1 −/− GES-1 and SULT2B1 −/− GES-1 cells transfected with Ad-SULT2B1b. f GES-1 and SULT2B1 −/− GES-1 cells were treated with 3-MCA (2 μg/mL) for 14 days. 3-MCA was added into the medium every 2–3 days. The cell morphology is presented. g The mRNA levels of pro- and anticancerous genes were detected by qRT-PCR in GES-1 and SULT2B1 −/− GES-1 cells in the absence or presence of 3-MCA treatment for 14 days. * P < 0.05, ** P < 0.01

Journal: Lipids in Health and Disease

Article Title: Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent

doi: 10.1186/s12944-019-1149-6

Figure Lengend Snippet: SULT2B1 deletion suppressed PI3K/AKT signaling in GES-1 cells and contributed to transformation upon 3-MCA induction. a , b The GES-1 and SULT2B1 −/− GES-1 cells were stimulated with IGF-1 (100 ng/mL, a ) or EGF (100 ng/mL, b ) for 5, 15, 30, 60, or 180 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. c GES-1 and SULT2B1 −/− GES-1 cells were stimulated with IGF-1 (100 ng/mL) or EGF (100 ng/mL) for 8 h. The mRNA levels of the genes involved in gastric epithelial function were measured by qRT-PCR. d Immunofluorescent staining of claudin-1 in GES-1 and SULT2B1 −/− GES-1 cells. The nuclei were revealed by DAPI staining (blue). e Wound healing assay of GES-1, SULT2B1 −/− GES-1 and SULT2B1 −/− GES-1 cells transfected with Ad-SULT2B1b. f GES-1 and SULT2B1 −/− GES-1 cells were treated with 3-MCA (2 μg/mL) for 14 days. 3-MCA was added into the medium every 2–3 days. The cell morphology is presented. g The mRNA levels of pro- and anticancerous genes were detected by qRT-PCR in GES-1 and SULT2B1 −/− GES-1 cells in the absence or presence of 3-MCA treatment for 14 days. * P < 0.05, ** P < 0.01

Article Snippet: The cells were transfected with human SULT2B1-specific siRNA duplex oligo ribonucleotides, with a 5′-GATCGAGATCATCTGCTTA-3′ targeting sequence, or negative control duplexes (RiboBio, Guangzhou, China) using Lipofectamine 3000.

Techniques: Transformation Assay, Western Blot, Quantitative RT-PCR, Staining, Wound Healing Assay, Transfection

24(R/S),25-EC and 27HC regulate gastric epithelial function. a The levels of 8 oxysterols (25HC, 27HC, 24(S) HC, 24(R/S),25-EC, 7αHC, 7βHC, 4βHC and 7KETO) were simultaneously detected by LC-MS in GES-1 and SULT2B1 −/− GES-1 cells with or without adenovirus-mediated SULT2B1b overexpression. b The GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27 HC for 12 h and then with IGF-1 (100 ng/mL) or EGF (100 ng/mL) for an additional 24 h. Cell proliferation was detected by the EdU incorporation assay. EdU fluorescence was normalized to that of Hoechst 33342. c , d GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27HC for 12 h and then with IGF-1 (100 ng/mL, c ) or EGF (100 ng/mL, d ) for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. e GES-1 cells were treated with 1 μmol/L 24(R/S), 25-EC and 27 HC for 12 h. The mRNA levels of the genes involved in gastric epithelial function were measured by qRT-PCR. f CCND1 and CCNA2 protein expression levels were detected by Western blotting. g GES-1 cells were treated with oxysterol (1 μmol/L) and 3-MCA (2 μg/mL) for 10 days. Oxysterol and 3-MCA were added into the medium every 2–3 days. The cell morphology is presented. The mRNA levels of pro- and anticancerous genes in the absence or presence of oxysterol/3-MCA treatment for 10 days were detected by qRT-PCR. * P < 0.05, ** P < 0.01

Journal: Lipids in Health and Disease

Article Title: Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent

doi: 10.1186/s12944-019-1149-6

Figure Lengend Snippet: 24(R/S),25-EC and 27HC regulate gastric epithelial function. a The levels of 8 oxysterols (25HC, 27HC, 24(S) HC, 24(R/S),25-EC, 7αHC, 7βHC, 4βHC and 7KETO) were simultaneously detected by LC-MS in GES-1 and SULT2B1 −/− GES-1 cells with or without adenovirus-mediated SULT2B1b overexpression. b The GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27 HC for 12 h and then with IGF-1 (100 ng/mL) or EGF (100 ng/mL) for an additional 24 h. Cell proliferation was detected by the EdU incorporation assay. EdU fluorescence was normalized to that of Hoechst 33342. c , d GES-1 cells were pretreated with 1 μmol/L of 24(R/S),25-EC and 27HC for 12 h and then with IGF-1 (100 ng/mL, c ) or EGF (100 ng/mL, d ) for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. e GES-1 cells were treated with 1 μmol/L 24(R/S), 25-EC and 27 HC for 12 h. The mRNA levels of the genes involved in gastric epithelial function were measured by qRT-PCR. f CCND1 and CCNA2 protein expression levels were detected by Western blotting. g GES-1 cells were treated with oxysterol (1 μmol/L) and 3-MCA (2 μg/mL) for 10 days. Oxysterol and 3-MCA were added into the medium every 2–3 days. The cell morphology is presented. The mRNA levels of pro- and anticancerous genes in the absence or presence of oxysterol/3-MCA treatment for 10 days were detected by qRT-PCR. * P < 0.05, ** P < 0.01

Article Snippet: The cells were transfected with human SULT2B1-specific siRNA duplex oligo ribonucleotides, with a 5′-GATCGAGATCATCTGCTTA-3′ targeting sequence, or negative control duplexes (RiboBio, Guangzhou, China) using Lipofectamine 3000.

Techniques: Liquid Chromatography with Mass Spectroscopy, Over Expression, Fluorescence, Western Blot, Quantitative RT-PCR, Expressing

SULT2B1 knockdown and oxysterols suppressed PI3K/AKT signaling in human primary stomach epithelial cells. Human primary stomach epithelial cells (HPSECs) were transfected with control-siRNA or SULT2B1-siRNA for 48 h. a The mRNA levels of genes involved in gastric epithelial function were measured by qRT-PCR. b Immunofluorescent staining of claudin-1 in HPSECs transfected with control-siRNA or SULT2B1-siRNA for 48 h. The nuclei were revealed by DAPI staining (blue). c The CCND1, CLND1 and CDH1 protein expression levels were detected by Western blotting. d After being transfected with control-siRNA or SULT2B1-siRNA for 48 h, the HPSECs were stimulated by EGF (100 ng/mL) for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. e The HPSECs were pretreated with 1 μmol/L of 24(R/S),25-EC or 27HC for 12 h and then treated with EGF (100 ng/mL for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. f HPSECs were treated with 1 μmol/L 24(R/S),25-EC and 27HC for 12 h. The CCND1, CLND1 and CDH1 protein expression levels were detected by Western blotting. * P < 0.05, ** P < 0.01

Journal: Lipids in Health and Disease

Article Title: Hydroxysteroid sulfotransferase 2B1 affects gastric epithelial function and carcinogenesis induced by a carcinogenic agent

doi: 10.1186/s12944-019-1149-6

Figure Lengend Snippet: SULT2B1 knockdown and oxysterols suppressed PI3K/AKT signaling in human primary stomach epithelial cells. Human primary stomach epithelial cells (HPSECs) were transfected with control-siRNA or SULT2B1-siRNA for 48 h. a The mRNA levels of genes involved in gastric epithelial function were measured by qRT-PCR. b Immunofluorescent staining of claudin-1 in HPSECs transfected with control-siRNA or SULT2B1-siRNA for 48 h. The nuclei were revealed by DAPI staining (blue). c The CCND1, CLND1 and CDH1 protein expression levels were detected by Western blotting. d After being transfected with control-siRNA or SULT2B1-siRNA for 48 h, the HPSECs were stimulated by EGF (100 ng/mL) for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. e The HPSECs were pretreated with 1 μmol/L of 24(R/S),25-EC or 27HC for 12 h and then treated with EGF (100 ng/mL for 20 or 90 min. The phosphorylation levels of AKT were detected by Western blotting. The p-AKT/t-AKT ratios are plotted. f HPSECs were treated with 1 μmol/L 24(R/S),25-EC and 27HC for 12 h. The CCND1, CLND1 and CDH1 protein expression levels were detected by Western blotting. * P < 0.05, ** P < 0.01

Article Snippet: The cells were transfected with human SULT2B1-specific siRNA duplex oligo ribonucleotides, with a 5′-GATCGAGATCATCTGCTTA-3′ targeting sequence, or negative control duplexes (RiboBio, Guangzhou, China) using Lipofectamine 3000.

Techniques: Transfection, Quantitative RT-PCR, Staining, Expressing, Western Blot

(A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates SULT2B1b KD. (C) Number of SULT2B1 reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: (A) Overview of the number of single cells sequenced. (B) Multidimensional scaling (MDS) plot highlighting the differences due to treatment between single cell groups. “C” indicates Control KD and “KD” indicates SULT2B1b KD. (C) Number of SULT2B1 reads indicating SULT2B1b expression at the single cell level is indicated for the Control KD (Control) or SULT2B1b KD (Knockdown) groups. The upper right-hand corner indicates the percentage of cells in each group with a zero read count. (D) Left: Violin plot representing expression (read count) of AR in scRNA-seq groups; Right: qRT-PCR expression of AR from bulk samples 48 hours after transfection. (E) Violin plots of the indicated AR target genes. Expression (read count) is indicated with adjusted p-values for each gene.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Expressing, Quantitative RT-PCR, Transfection

Violin plots representing expression (read count) of indicated NF-κB target genes in Control KD versus SULT2B1b KD groups of the scRNA-seq analysis, with adjusted p-value indicated in the upper left-hand corner.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: Violin plots representing expression (read count) of indicated NF-κB target genes in Control KD versus SULT2B1b KD groups of the scRNA-seq analysis, with adjusted p-value indicated in the upper left-hand corner.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Expressing

LNCaP and C4-2 cells were transfected with control or SULT2B1b siRNA and evaluated for (A-B) TNF expression or (C-D) NF-κB activity. (A) qRT-PCR for TNF expression 72 hours after siRNA KD. (B) TNF protein expression was quantified by cell lysate-based ELISA 72 hours after siRNA KD. (C) Luciferase assay completed 48 hours after siRNA transfection. NF-κB luciferase activity was normalized to Renilla luciferase. Samples were further normalized to control wells without siRNA transfection. Bars indicate the mean +/− SEM of at least four independent experiments using a Student’s t-test. (D) Western blot of indicated proteins 72 hours after siRNA KD.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: LNCaP and C4-2 cells were transfected with control or SULT2B1b siRNA and evaluated for (A-B) TNF expression or (C-D) NF-κB activity. (A) qRT-PCR for TNF expression 72 hours after siRNA KD. (B) TNF protein expression was quantified by cell lysate-based ELISA 72 hours after siRNA KD. (C) Luciferase assay completed 48 hours after siRNA transfection. NF-κB luciferase activity was normalized to Renilla luciferase. Samples were further normalized to control wells without siRNA transfection. Bars indicate the mean +/− SEM of at least four independent experiments using a Student’s t-test. (D) Western blot of indicated proteins 72 hours after siRNA KD.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Transfection, Expressing, Activity Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Luciferase, Western Blot

(A) pRT-PCR indicates SULT2B1b and TNF expression in LNCaP cells that were consecutively transfected with SULT2B1b siRNA followed by TNF siRNA. Significance was determined with a one-way ANOVA with a multiple comparisons test. (B) NF-κB luciferase assay was completed 48 hours after siRNA transfection and relative luciferase units (RLU) are shown. Bars indicate the mean +/− SEM of at least triplicate samples by two-way ANOVA considering only Control KD and SULT2B1b KD groups and the graph is representative of two independent experiments. (C) LNCaP cells were pre-treated with TNF neutralizing antibody (40 µg/mL) and then transfected with Control or SULT2B1b siRNA or treated with indicated concentrations of exogenous TNFα. Approximately 72 hours post-siRNA transfection, cells were harvested and prepared for cell cycle analysis by flow cytometry. Bars indicate the average percent sub-G1 nuclei +/− SEM of triplicate samples. This graph is representative of at least two independent experiments.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: (A) pRT-PCR indicates SULT2B1b and TNF expression in LNCaP cells that were consecutively transfected with SULT2B1b siRNA followed by TNF siRNA. Significance was determined with a one-way ANOVA with a multiple comparisons test. (B) NF-κB luciferase assay was completed 48 hours after siRNA transfection and relative luciferase units (RLU) are shown. Bars indicate the mean +/− SEM of at least triplicate samples by two-way ANOVA considering only Control KD and SULT2B1b KD groups and the graph is representative of two independent experiments. (C) LNCaP cells were pre-treated with TNF neutralizing antibody (40 µg/mL) and then transfected with Control or SULT2B1b siRNA or treated with indicated concentrations of exogenous TNFα. Approximately 72 hours post-siRNA transfection, cells were harvested and prepared for cell cycle analysis by flow cytometry. Bars indicate the average percent sub-G1 nuclei +/− SEM of triplicate samples. This graph is representative of at least two independent experiments.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Expressing, Transfection, Luciferase, Cell Cycle Assay, Flow Cytometry

LNCaP (A) or C4-2 (B) cells were transfected with Control or SULT2B1b siRNA prior to addition of indicated concentrations of TNF. All cells were harvested 72 hours later and prepared for cell cycle analysis by flow cytometry. Bars indicate the average percent sub-G1 nuclei +/− SEM of triplicate wells. For all TNF-treated cells in (A) and (B), SULT2B1b KD significantly enhances sub-G1 nuclei percentage over Control KD cells in both LNCaP and C4-2 cells by two-way ANOVA (p<0.0001). Graphs are representative of three independent experiments. (C-D) SULT2B1b expression was evaluated by (C) qRT-PCR and (D) western blot in LNCaP-SULT2B1b cells. (E) LNCaP-SULT2B1b cells were treated with doxycycline prior to addition of indicated concentrations of TNF. After 72 hours, all cells were harvested and prepared for cell cycle analysis by flow cytometry. Bars indicate the mean of % sub-G1 nuclei +/− SEM of triplicate wells evaluated by two-way ANOVA with multiple comparisons test. The graph is representative of duplicate experiments.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: LNCaP (A) or C4-2 (B) cells were transfected with Control or SULT2B1b siRNA prior to addition of indicated concentrations of TNF. All cells were harvested 72 hours later and prepared for cell cycle analysis by flow cytometry. Bars indicate the average percent sub-G1 nuclei +/− SEM of triplicate wells. For all TNF-treated cells in (A) and (B), SULT2B1b KD significantly enhances sub-G1 nuclei percentage over Control KD cells in both LNCaP and C4-2 cells by two-way ANOVA (p<0.0001). Graphs are representative of three independent experiments. (C-D) SULT2B1b expression was evaluated by (C) qRT-PCR and (D) western blot in LNCaP-SULT2B1b cells. (E) LNCaP-SULT2B1b cells were treated with doxycycline prior to addition of indicated concentrations of TNF. After 72 hours, all cells were harvested and prepared for cell cycle analysis by flow cytometry. Bars indicate the mean of % sub-G1 nuclei +/− SEM of triplicate wells evaluated by two-way ANOVA with multiple comparisons test. The graph is representative of duplicate experiments.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Transfection, Cell Cycle Assay, Flow Cytometry, Expressing, Quantitative RT-PCR, Western Blot

(A) Gene co-expression indicates SULT2B1 negatively correlates with TNF, CD40LG, TRADD, and TRAF2 in bone marrow-derived metastatic prostate cancer samples from individuals with no prior treatment. (B) Co-expression of SULT2B1 with NFKB1, FADD, and DR4 determines significant negative correlations with each of these genes in lymph node-derived CRPC samples. (C) Gene co-expression of SULT2B1 with RANKL demonstrates a strong positive correlation in bone marrow-derived prostate cancer samples. Pearson correlation coefficients (r) and p-values determined by a permutation test (p) are indicated for each correlation plot. Dots indicate samples obtained from individual patients.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: (A) Gene co-expression indicates SULT2B1 negatively correlates with TNF, CD40LG, TRADD, and TRAF2 in bone marrow-derived metastatic prostate cancer samples from individuals with no prior treatment. (B) Co-expression of SULT2B1 with NFKB1, FADD, and DR4 determines significant negative correlations with each of these genes in lymph node-derived CRPC samples. (C) Gene co-expression of SULT2B1 with RANKL demonstrates a strong positive correlation in bone marrow-derived prostate cancer samples. Pearson correlation coefficients (r) and p-values determined by a permutation test (p) are indicated for each correlation plot. Dots indicate samples obtained from individual patients.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Expressing, Derivative Assay

Diagram of proposed SULT2B1b influence on death receptor signaling and AR activity in advanced, hormone-naive prostate cancer and CRPC cells.

Journal: Molecular cancer research : MCR

Article Title: Cholesterol Sulfotransferase SULT2B1b Modulates Sensitivity to Death Receptor Ligand TNF alpha in Castration Resistant Prostate Cancer

doi: 10.1158/1541-7786.MCR-18-1054

Figure Lengend Snippet: Diagram of proposed SULT2B1b influence on death receptor signaling and AR activity in advanced, hormone-naive prostate cancer and CRPC cells.

Article Snippet: Since scRNA-seq requires viable cells, LNCaP were harvested 48 hours after non-targeting or SULT2B1 siRNA (Control KD or SULT2B1b KD, respectively) transfection and then subjected to viable cell sorting prior to single-cell isolation on the Fluidigm C1 Single-Cell Auto Prep System ( Supplementary Figure 1A ).

Techniques: Activity Assay