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    ATCC sw13 human adenocarcinoma cells
    Growth of E protein 390 mutants in cell culture. Vero, <t>SW13,</t> and C6/36 cells were infected with MVE-1-51, the Asp 390 control virus, or 1 of 10 E protein 390 mutants at multiplicity of ∼0.1, determined for each cell type. Unbound virus was removed after 1 h of adsorption, and growth medium was added. At 16, 20, 24, and 28 hpi, supernatants were collected from Vero and SW13 cells for titration of virus infectivity, in the respective cell types. For growth assays in C6/36 cells, supernatant samples were taken at 24, 28, and 44 hpi, and virus titers were determined as focus-forming units (FFU) by immunofluorescence in C6/36 cells. Results for Vero and SW13 growth assays are from two separate experiments (shown in different graphs).
    Sw13 Human Adenocarcinoma Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sw13 human adenocarcinoma cells/product/ATCC
    Average 95 stars, based on 67 article reviews
    Price from $9.99 to $1999.99
    sw13 human adenocarcinoma cells - by Bioz Stars, 2020-07
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    93
    Millipore sw13 human adrenocortical cells
    (A) Aldosterone production in human CAR47, <t>SW13,</t> and various NCI-H295-derived cell models including HAC cell lines. All cells were incubated for 48 h followed by aldosterone measurement in the experimental medium. Results represent a minimum of 3 independent
    Sw13 Human Adrenocortical Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sw13 human adrenocortical cells/product/Millipore
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    sw13 human adrenocortical cells - by Bioz Stars, 2020-07
    93/100 stars
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    Image Search Results


    Growth of E protein 390 mutants in cell culture. Vero, SW13, and C6/36 cells were infected with MVE-1-51, the Asp 390 control virus, or 1 of 10 E protein 390 mutants at multiplicity of ∼0.1, determined for each cell type. Unbound virus was removed after 1 h of adsorption, and growth medium was added. At 16, 20, 24, and 28 hpi, supernatants were collected from Vero and SW13 cells for titration of virus infectivity, in the respective cell types. For growth assays in C6/36 cells, supernatant samples were taken at 24, 28, and 44 hpi, and virus titers were determined as focus-forming units (FFU) by immunofluorescence in C6/36 cells. Results for Vero and SW13 growth assays are from two separate experiments (shown in different graphs).

    Journal: Journal of Virology

    Article Title: Substitutions at the Putative Receptor-Binding Site of an Encephalitic Flavivirus Alter Virulence and Host Cell Tropism and Reveal a Role for Glycosaminoglycans in Entry

    doi:

    Figure Lengend Snippet: Growth of E protein 390 mutants in cell culture. Vero, SW13, and C6/36 cells were infected with MVE-1-51, the Asp 390 control virus, or 1 of 10 E protein 390 mutants at multiplicity of ∼0.1, determined for each cell type. Unbound virus was removed after 1 h of adsorption, and growth medium was added. At 16, 20, 24, and 28 hpi, supernatants were collected from Vero and SW13 cells for titration of virus infectivity, in the respective cell types. For growth assays in C6/36 cells, supernatant samples were taken at 24, 28, and 44 hpi, and virus titers were determined as focus-forming units (FFU) by immunofluorescence in C6/36 cells. Results for Vero and SW13 growth assays are from two separate experiments (shown in different graphs).

    Article Snippet: Vero (African Green monkey kidney), BHK-21 (baby hamster kidney), and SW13 (human adenocarcinoma) cells were all from the American Type Culture Collection (Bethesda, Md.) and maintained in Eagle's minimal essential medium (EMEM) plus nonessential amino acids and 5% fetal calf serum (FCS).

    Techniques: Cell Culture, Infection, Adsorption, Titration, Immunofluorescence

    Uptake of MVE into SW13 cells. The kinetics of uptake of the Asp 390 control and His 390 mutant viruses into SW13 cells was determined as described in Materials and Methods. Percent uptake for each time point is expressed as the ratio of the number of plaques obtained after acid treatment and the number of plaques obtained on a control monolayer in the absence of acid treatment.

    Journal: Journal of Virology

    Article Title: Substitutions at the Putative Receptor-Binding Site of an Encephalitic Flavivirus Alter Virulence and Host Cell Tropism and Reveal a Role for Glycosaminoglycans in Entry

    doi:

    Figure Lengend Snippet: Uptake of MVE into SW13 cells. The kinetics of uptake of the Asp 390 control and His 390 mutant viruses into SW13 cells was determined as described in Materials and Methods. Percent uptake for each time point is expressed as the ratio of the number of plaques obtained after acid treatment and the number of plaques obtained on a control monolayer in the absence of acid treatment.

    Article Snippet: Vero (African Green monkey kidney), BHK-21 (baby hamster kidney), and SW13 (human adenocarcinoma) cells were all from the American Type Culture Collection (Bethesda, Md.) and maintained in Eagle's minimal essential medium (EMEM) plus nonessential amino acids and 5% fetal calf serum (FCS).

    Techniques: Mutagenesis

    Inhibition by heparin of virus infectivity in Vero, SW13, and BHK cells. (A) The Asp 390 control and His 390 , Gly 390 , and Lys 390 mutant viruses (200 PFU) were incubated with heparin (0, 50, 100, and 200 μg/ml) prior to their addition to cells pretreated with HBSS-BSA containing heparin at 0, 50, 100, or 200 μg/ml. Agar overlay was added to allow plaque formation after 1 h of adsorption at 37°C. Inhibition by heparin was calculated using the formula [(plaque number on nontreated cells − plaque number on heparin-treated cells)/(plaque number from non-treated cells)] × 100. (B) Inhibition of infectivity by heparin (200 μg/ml) of 10 E protein 390 mutants was assayed in parallel with the Asp 390 control virus as in panel A. The percent inhibition by heparin of the Asp 390 virus is subtracted from that for the E protein 390 mutants obtained in the same experiment. Results (mean and standard error) from three independent experiments are shown for each mutant. (C) Inhibition by heparin of virus binding to Vero and SW13 cells. The Asp 390 control virus and the His 390 mutant were incubated with heparin (0, 20, 50, 100, or 200 μg/ml) for 15 min at 4°C and then added to chilled Vero or SW13 cells treated with heparin (0 to 200 μg/ml). After 1 h of adsorption at 4°C, cell monolayers were washed, and agar overlay was added to allow plaque development. Percent inhibition by heparin is calculated as in panel A. Results (mean and standard error) from two independent experiments are shown.

    Journal: Journal of Virology

    Article Title: Substitutions at the Putative Receptor-Binding Site of an Encephalitic Flavivirus Alter Virulence and Host Cell Tropism and Reveal a Role for Glycosaminoglycans in Entry

    doi:

    Figure Lengend Snippet: Inhibition by heparin of virus infectivity in Vero, SW13, and BHK cells. (A) The Asp 390 control and His 390 , Gly 390 , and Lys 390 mutant viruses (200 PFU) were incubated with heparin (0, 50, 100, and 200 μg/ml) prior to their addition to cells pretreated with HBSS-BSA containing heparin at 0, 50, 100, or 200 μg/ml. Agar overlay was added to allow plaque formation after 1 h of adsorption at 37°C. Inhibition by heparin was calculated using the formula [(plaque number on nontreated cells − plaque number on heparin-treated cells)/(plaque number from non-treated cells)] × 100. (B) Inhibition of infectivity by heparin (200 μg/ml) of 10 E protein 390 mutants was assayed in parallel with the Asp 390 control virus as in panel A. The percent inhibition by heparin of the Asp 390 virus is subtracted from that for the E protein 390 mutants obtained in the same experiment. Results (mean and standard error) from three independent experiments are shown for each mutant. (C) Inhibition by heparin of virus binding to Vero and SW13 cells. The Asp 390 control virus and the His 390 mutant were incubated with heparin (0, 20, 50, 100, or 200 μg/ml) for 15 min at 4°C and then added to chilled Vero or SW13 cells treated with heparin (0 to 200 μg/ml). After 1 h of adsorption at 4°C, cell monolayers were washed, and agar overlay was added to allow plaque development. Percent inhibition by heparin is calculated as in panel A. Results (mean and standard error) from two independent experiments are shown.

    Article Snippet: Vero (African Green monkey kidney), BHK-21 (baby hamster kidney), and SW13 (human adenocarcinoma) cells were all from the American Type Culture Collection (Bethesda, Md.) and maintained in Eagle's minimal essential medium (EMEM) plus nonessential amino acids and 5% fetal calf serum (FCS).

    Techniques: Inhibition, Infection, Mutagenesis, Incubation, Adsorption, Binding Assay

    Details of KF network formation in living SW13 cells synthesizing HK8-CFP and HK18-YFP fluorescent hybrids. (A) Epifluorescence microscopy (detection of HK18-YFP) of a characteristic peripheral segment (position of plasma membrane indicated at 3.5 min) highlighting intermediate steps of KF network production. Two KF precursors are marked by color and followed until integration into the filament system. Note the formation of small granules below the cell surface that elongate, fuse, and become part of the KF network. The corresponding movie 5 shows further details. (B) Size changes of a single particle that was tracked from its first detection until integration into the network. For clarity's sake only the pixels associated with that particular particle are shown. Sampling frequency, 30 s. (C) The graph is derived from B, showing a surface view of the altering particle dimensions demonstrating a continuous, nonincremental particle elongation (l) before fusion with the KF network over time (t). (D) 3D reconstruction (voltex presentation) of the HK18-YFP fluorescence as recorded by confocal laser scanning microscopy of a peripheral region revealing spatial characteristics of various intermediates of filament assembly in relationship to intracellular topology (position of cell border demarcated). The volume of the top segment has the dimensions 20 × 20 × 3.5 μm. The bottom cube is a cross section to specifically highlight the increase in diameter of KF precursors only after integration into the peripheral KF network. (E) Plot summarizing the quantitative analysis of 10 time-space diagrams (see, e.g., C) depicting the increase in length (top graph) and diameter (bottom graph). Note the continuous elongation of particles during the entire 20 min in contrast to the increase in diameter that is limited to the first 5 min. Bars, SD. Bars, 1 μm in A and D.

    Journal: Molecular Biology of the Cell

    Article Title: Identification of Novel Principles of Keratin Filament Network Turnover in Living Cells V⃞

    doi: 10.1091/mbc.E03-09-0707

    Figure Lengend Snippet: Details of KF network formation in living SW13 cells synthesizing HK8-CFP and HK18-YFP fluorescent hybrids. (A) Epifluorescence microscopy (detection of HK18-YFP) of a characteristic peripheral segment (position of plasma membrane indicated at 3.5 min) highlighting intermediate steps of KF network production. Two KF precursors are marked by color and followed until integration into the filament system. Note the formation of small granules below the cell surface that elongate, fuse, and become part of the KF network. The corresponding movie 5 shows further details. (B) Size changes of a single particle that was tracked from its first detection until integration into the network. For clarity's sake only the pixels associated with that particular particle are shown. Sampling frequency, 30 s. (C) The graph is derived from B, showing a surface view of the altering particle dimensions demonstrating a continuous, nonincremental particle elongation (l) before fusion with the KF network over time (t). (D) 3D reconstruction (voltex presentation) of the HK18-YFP fluorescence as recorded by confocal laser scanning microscopy of a peripheral region revealing spatial characteristics of various intermediates of filament assembly in relationship to intracellular topology (position of cell border demarcated). The volume of the top segment has the dimensions 20 × 20 × 3.5 μm. The bottom cube is a cross section to specifically highlight the increase in diameter of KF precursors only after integration into the peripheral KF network. (E) Plot summarizing the quantitative analysis of 10 time-space diagrams (see, e.g., C) depicting the increase in length (top graph) and diameter (bottom graph). Note the continuous elongation of particles during the entire 20 min in contrast to the increase in diameter that is limited to the first 5 min. Bars, SD. Bars, 1 μm in A and D.

    Article Snippet: The following cell lines were used and cultured according to the specifications given by ATCC or the respective references: rat kangaroo kidney PtK2 cells (ATCC CCL-56), Madin-Darby canine kidney (MDCK) cells (clone 20; ATCC CCL-34), bovine kidney-derived MDBK cells (ATCC CCL-22), human mammary adenocarcinoma-derived MCF7 cells (ATCC HTB-22), human hepatocellular carcinoma-derived PLC cells (ATCC CRL8024), human colon carcinoma CaCo-2 cells (ATCC HTB-37), human immortalized HaCaT keratinocytes , and human small-cell carcinoma SW13 cells derived from the adrenal cortex (ATCC CCL-105).

    Techniques: Epifluorescence Microscopy, Sampling, Derivative Assay, Fluorescence, Confocal Laser Scanning Microscopy

    Inhibitors of HDAC1 increase BRM and VIM expression in a time-dependent manner. Total RNA was isolated from untreated SW13- cells (0 h), and SW13- cells treated with 0.15 μM MGCD0103, 0.51 μM MS-275, or 2 nM FK228 for 24, 48, and 72 h. Fold change in ( a ) BRM and ( b ) VIM mRNA expression from 0 h was determined by real-time PCR analysis using the 2 -ΔΔCt method normalized to GAPDH. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: Inhibitors of HDAC1 increase BRM and VIM expression in a time-dependent manner. Total RNA was isolated from untreated SW13- cells (0 h), and SW13- cells treated with 0.15 μM MGCD0103, 0.51 μM MS-275, or 2 nM FK228 for 24, 48, and 72 h. Fold change in ( a ) BRM and ( b ) VIM mRNA expression from 0 h was determined by real-time PCR analysis using the 2 -ΔΔCt method normalized to GAPDH. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Expressing, Isolation, Mass Spectrometry, Real-time Polymerase Chain Reaction

    Validation of select genes related to epigenetic chromatin modification and remodeling by qPCR. Relative expression of a total of bromodomain containing 2 (BRD2), HDAC7, HDAC9, HDAC10, lysine (K)-specific demethylase 5c (KDM5C), and methyl-CpG binding domain protein 2 (MBD2) was examined in stable SW13- and SW13+ subtypes and SW13- cells which had been treated for 24 h with either 0.51 μM MS-275 or 2 nM FK228. Fold-regulation was determined by the 2 -ΔΔCT method using GAPDH as the invariant control. Data are presented as mean ± SEM. *Denotes statistical difference from SW13-, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: Validation of select genes related to epigenetic chromatin modification and remodeling by qPCR. Relative expression of a total of bromodomain containing 2 (BRD2), HDAC7, HDAC9, HDAC10, lysine (K)-specific demethylase 5c (KDM5C), and methyl-CpG binding domain protein 2 (MBD2) was examined in stable SW13- and SW13+ subtypes and SW13- cells which had been treated for 24 h with either 0.51 μM MS-275 or 2 nM FK228. Fold-regulation was determined by the 2 -ΔΔCT method using GAPDH as the invariant control. Data are presented as mean ± SEM. *Denotes statistical difference from SW13-, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Modification, Real-time Polymerase Chain Reaction, Expressing, Binding Assay, Mass Spectrometry

    HDAC1 inhibitors reduce cell proliferation and promote restoration of VIM and BRM protein to varying degrees. a SW13- and SW13+ cells as well as SW13- cells treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h were labeled with EdU for 24 h to determine proliferation rates. b Representative images of two independent experiments performed in triplicate are shown. c Nuclear protein was isolated from SW13- and SW13+ cells, as well as from SW13- cells which had been treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h and the expression of SWI/SNF protein components were examined by western blot. Total histone H3 was used as a loading control. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: HDAC1 inhibitors reduce cell proliferation and promote restoration of VIM and BRM protein to varying degrees. a SW13- and SW13+ cells as well as SW13- cells treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h were labeled with EdU for 24 h to determine proliferation rates. b Representative images of two independent experiments performed in triplicate are shown. c Nuclear protein was isolated from SW13- and SW13+ cells, as well as from SW13- cells which had been treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h and the expression of SWI/SNF protein components were examined by western blot. Total histone H3 was used as a loading control. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Mass Spectrometry, Labeling, Isolation, Expressing, Western Blot

    Histone acetylation in stable SW13- and SW13+ subtypes and following treatment with the HDAC inhibitors MS-275 and FK228. Pure histones were isolated from SW13- and SW13+ cells, as well as from SW13- cells which had been treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h. Histones were separated on tris-tricine gels and transferred to a PVDF membrane for blotting and assessment of histone modifications. Total histone H3 was used as a loading control. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: Histone acetylation in stable SW13- and SW13+ subtypes and following treatment with the HDAC inhibitors MS-275 and FK228. Pure histones were isolated from SW13- and SW13+ cells, as well as from SW13- cells which had been treated with either 0.51 μM MS-275 or 2 nM FK228 for 24 h. Histones were separated on tris-tricine gels and transferred to a PVDF membrane for blotting and assessment of histone modifications. Total histone H3 was used as a loading control. Data are presented as mean ± SEM. Superscripts indicate statistical significance, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Mass Spectrometry, Isolation

    SW13- and SW13+ subtype characterization: differences in morphology, actin organization, vimentin expression, and VIM and BRM levels. a Subtypes have distinct morphology, actin organization, and levels of vimentin expression. Left-hand panel: light microscopy photographs; Middle panel: visualization of actin filaments with fluorescent phalloidin; Right-hand panel: expression of vimentin by immunofluorescence. Images were taken using a 40× oil-immersion objective lens. b qPCR reveals VIM and BRM mRNA expression is ~ 8 -fold higher in the SW13+ cells compared to the SW13- cells. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: SW13- and SW13+ subtype characterization: differences in morphology, actin organization, vimentin expression, and VIM and BRM levels. a Subtypes have distinct morphology, actin organization, and levels of vimentin expression. Left-hand panel: light microscopy photographs; Middle panel: visualization of actin filaments with fluorescent phalloidin; Right-hand panel: expression of vimentin by immunofluorescence. Images were taken using a 40× oil-immersion objective lens. b qPCR reveals VIM and BRM mRNA expression is ~ 8 -fold higher in the SW13+ cells compared to the SW13- cells. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Expressing, Light Microscopy, Immunofluorescence, Real-time Polymerase Chain Reaction

    SW13- cells are more proliferative and have higher rates of anchorage independent growth than SW13+. SW13- and SW13+ cells were seeded in 6-well plates at 1 × 10 4 cells per well and counted using a hemocytometer and trypan exclusion every 24 h for 7 days. a The growth rate of SW13- cells is significantly higher than SW13+ cells. b SW13- cells have higher rates of proliferation as determined by the higher number of EdU positive cells ( green ) per total number of cells indicated by Hoechst 33342 staining ( blue ). Images were taken using a 10× objective lens and quantitated ( f ) using ImageJ software. c Soft agar colony formation assays revealed SW13- cells exhibit increased rates of anchorage independent growth as indicated by ( d ) increased colony number and ( e ) increased colony size. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: SW13- cells are more proliferative and have higher rates of anchorage independent growth than SW13+. SW13- and SW13+ cells were seeded in 6-well plates at 1 × 10 4 cells per well and counted using a hemocytometer and trypan exclusion every 24 h for 7 days. a The growth rate of SW13- cells is significantly higher than SW13+ cells. b SW13- cells have higher rates of proliferation as determined by the higher number of EdU positive cells ( green ) per total number of cells indicated by Hoechst 33342 staining ( blue ). Images were taken using a 10× objective lens and quantitated ( f ) using ImageJ software. c Soft agar colony formation assays revealed SW13- cells exhibit increased rates of anchorage independent growth as indicated by ( d ) increased colony number and ( e ) increased colony size. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Staining, Software

    The ability of selected HDAC inhibitors to induce the switching from the SW13- to the SW13+ subtype. a Induction of vimentin expression was assayed at 24, 48, and 72 h using fluorescence intensity of vimentin normalized to DAPI staining at each time point ( left columns ). Dose response was measured by fold induction in BRM mRNA using qPCR for two doses of each inhibitor ( right columns ). b Representative immunofluorescence images which illustrate the range of vimentin induction in panel ( a ). Scale ranges from no detectable vimentin immunofluorescence signal (−) to + representing

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: The ability of selected HDAC inhibitors to induce the switching from the SW13- to the SW13+ subtype. a Induction of vimentin expression was assayed at 24, 48, and 72 h using fluorescence intensity of vimentin normalized to DAPI staining at each time point ( left columns ). Dose response was measured by fold induction in BRM mRNA using qPCR for two doses of each inhibitor ( right columns ). b Representative immunofluorescence images which illustrate the range of vimentin induction in panel ( a ). Scale ranges from no detectable vimentin immunofluorescence signal (−) to + representing

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Expressing, Fluorescence, Staining, Real-time Polymerase Chain Reaction, Immunofluorescence

    SW13+ cells display properties associated with increased rates of metastasis to a greater extent than do SW13- cells. a and c Transwell migration assays indicate SW13+ cells have increased rates of chemotaxis compared to the SW13- cells at 24 and 48 h. Images were taken using a 4× objective lens ( b and d ) SW13+ cells exhibited higher levels of MMP2/9 (collagenase) activity as measured by increased fluorescence after incubation with a fluorescein-quenched gelatin substrate (DQ gelatin). Addition of a broad-spectrum MMP inhibitor reduced the fluorescence signal in both subtypes. Images were taken using a 40× objective lens. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Journal: BMC Cancer

    Article Title: Epigenetically maintained SW13+ and SW13- subtypes have different oncogenic potential and convert with HDAC1 inhibition

    doi: 10.1186/s12885-016-2353-7

    Figure Lengend Snippet: SW13+ cells display properties associated with increased rates of metastasis to a greater extent than do SW13- cells. a and c Transwell migration assays indicate SW13+ cells have increased rates of chemotaxis compared to the SW13- cells at 24 and 48 h. Images were taken using a 4× objective lens ( b and d ) SW13+ cells exhibited higher levels of MMP2/9 (collagenase) activity as measured by increased fluorescence after incubation with a fluorescein-quenched gelatin substrate (DQ gelatin). Addition of a broad-spectrum MMP inhibitor reduced the fluorescence signal in both subtypes. Images were taken using a 40× objective lens. Data are presented as mean ± SEM. *Denotes statistical difference between subtypes, p

    Article Snippet: Cell culture SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10 % fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Migration, Chemotaxis Assay, Activity Assay, Fluorescence, Incubation

    Expression of peripherin isoforms in transfected SW13 vim( − ) cells. (A) Mammalian expression plasmids encoding the peripherin isoforms, Per 58, Per 56, or Per 61, and also the full-length peripherin gene (Per gene) were transfected into SW13 vim(−) cells and harvested 48 h later. Equal protein loadings (10 μg) of harvested cells were resolved on 7.5% (wt/vol) SDS–polyacrylamide gels, and then transferred to PVDF membrane and probed with a monoclonal antibody recognizing peripherin (MAB1527). For Per 61, immunoprecipitation using peripherin polyclonal antiserum (AB1530) was used to obtain sufficient quantities for detection on immunoblots. The immunoblot shows that expression of the plasmids encoding Per 58, Per 56, and Per 61, as indicated, generate proteins of the expected molecular masses with the position of the 62-kD marker shown (arrow). The major peripherin species expressed from the full-length peripherin gene (Per gene) corresponds to the molecular mass of isoform Per 58. (B) SW13 vim(−) cells transfected with the individual peripherin isoform expression plasmids were labeled immunocytochemically with monoclonal antibody to peripherin (MAB1527) to ascertain the cytoplasmic distribution of the expressed proteins. From the figure it can be seen that both Per 58 and Per 56 were capable of self-assembly to form an IF network that was distributed evenly throughout the cytoplasm. These results are similar to those obtained with the full-length mouse peripherin gene ( Beaulieu et al., 1999b ). In contrast, Per 61 was assembly incompetent, forming aggregates of uneven size and distribution. Bar, 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: A neurotoxic peripherin splice variant in a mouse model of ALS

    doi: 10.1083/jcb.200205027

    Figure Lengend Snippet: Expression of peripherin isoforms in transfected SW13 vim( − ) cells. (A) Mammalian expression plasmids encoding the peripherin isoforms, Per 58, Per 56, or Per 61, and also the full-length peripherin gene (Per gene) were transfected into SW13 vim(−) cells and harvested 48 h later. Equal protein loadings (10 μg) of harvested cells were resolved on 7.5% (wt/vol) SDS–polyacrylamide gels, and then transferred to PVDF membrane and probed with a monoclonal antibody recognizing peripherin (MAB1527). For Per 61, immunoprecipitation using peripherin polyclonal antiserum (AB1530) was used to obtain sufficient quantities for detection on immunoblots. The immunoblot shows that expression of the plasmids encoding Per 58, Per 56, and Per 61, as indicated, generate proteins of the expected molecular masses with the position of the 62-kD marker shown (arrow). The major peripherin species expressed from the full-length peripherin gene (Per gene) corresponds to the molecular mass of isoform Per 58. (B) SW13 vim(−) cells transfected with the individual peripherin isoform expression plasmids were labeled immunocytochemically with monoclonal antibody to peripherin (MAB1527) to ascertain the cytoplasmic distribution of the expressed proteins. From the figure it can be seen that both Per 58 and Per 56 were capable of self-assembly to form an IF network that was distributed evenly throughout the cytoplasm. These results are similar to those obtained with the full-length mouse peripherin gene ( Beaulieu et al., 1999b ). In contrast, Per 61 was assembly incompetent, forming aggregates of uneven size and distribution. Bar, 10 μm.

    Article Snippet: Transient transfections A human adrenal carcinoma cell line, SW13 vim(−), that lacks cytoplasmic IFs, was derived by dilutional cloning of SW13 cells obtained from the American Type Culture Collection.

    Techniques: Expressing, Transfection, Immunoprecipitation, Western Blot, Marker, Labeling

    Characterization of specific antisera to peripherin isoforms. (A–F) SW13 vim(−) cells transfected with plasmid expression vectors encoding Per 58, Per 56, or Per 61 were labeled by indirect immunofluorescence with anti–Per 56 or anti–Per 61. There was no labeling of Per 58 by either antibody (A and B). Both anti–Per 56 and anti–Per 61 were specific for their respective proteins, Per 56 appearing filamentous (D) and Per 61 speckled (E). Bars: (A–D and F) 20 μm; (E) 30 μm. (G) Immunblots of cell extracts from NIH 3T3 cells transfected with Per 58 (lane 1), Per 56 (lane 2), or Per 61 (lane 3) were probed with polyclonal antiserum to peripherin (Poly Per; AB1530); anti–Per 61 and anti–Per 56. The results obtained confirmed the specificity of the Per 56 and Per 61 antisera.

    Journal: The Journal of Cell Biology

    Article Title: A neurotoxic peripherin splice variant in a mouse model of ALS

    doi: 10.1083/jcb.200205027

    Figure Lengend Snippet: Characterization of specific antisera to peripherin isoforms. (A–F) SW13 vim(−) cells transfected with plasmid expression vectors encoding Per 58, Per 56, or Per 61 were labeled by indirect immunofluorescence with anti–Per 56 or anti–Per 61. There was no labeling of Per 58 by either antibody (A and B). Both anti–Per 56 and anti–Per 61 were specific for their respective proteins, Per 56 appearing filamentous (D) and Per 61 speckled (E). Bars: (A–D and F) 20 μm; (E) 30 μm. (G) Immunblots of cell extracts from NIH 3T3 cells transfected with Per 58 (lane 1), Per 56 (lane 2), or Per 61 (lane 3) were probed with polyclonal antiserum to peripherin (Poly Per; AB1530); anti–Per 61 and anti–Per 56. The results obtained confirmed the specificity of the Per 56 and Per 61 antisera.

    Article Snippet: Transient transfections A human adrenal carcinoma cell line, SW13 vim(−), that lacks cytoplasmic IFs, was derived by dilutional cloning of SW13 cells obtained from the American Type Culture Collection.

    Techniques: Transfection, Plasmid Preparation, Expressing, Labeling, Immunofluorescence

    Assembly characteristics of peripherin isoforms in transfected SW13 vim( − ) cells. SW13 vim(−) cells were transfected with expression plasmids encoding the different peripherin isoforms together with plasmids encoding NF-L, NF-M, or NF-H. Assembly properties of the coexpressed proteins were analyzed by double immunofluorescence labeling using monoclonal antibody to peripherin (MAB1527; red) and polyclonal antiserum (green) to NF-L (AB1983), NF-M (AB1981), and NF-H (AB 1982) in the respective cotransfected cells. Isoforms Per 58 and Per 56 were capable of coassembly with all three neurofilament subunits (A, A′; B, B′; D, D′; E, E′; I, I′; and J, J′), whereas isoform Per 61 was incapable of coassembly into an IF network with any of the neurofilament subunits (C, C′; H, H′; and O, O′). Additional phenotypes were observed in cells cotransfected with Per 58 or Per 56 and NF-M or NF-H. These were filamentous bundles of peripherin in the presence of nonassembled NF-M (F, F′ and G, G′) or NF-H (K, K′ and L, L′; arrows in K indicate filaments) and nonassembled Per 58 or Per 56 in the presence of nonassembled NF-H (M, M′ and N, N′). Bar, 14 μm.

    Journal: The Journal of Cell Biology

    Article Title: A neurotoxic peripherin splice variant in a mouse model of ALS

    doi: 10.1083/jcb.200205027

    Figure Lengend Snippet: Assembly characteristics of peripherin isoforms in transfected SW13 vim( − ) cells. SW13 vim(−) cells were transfected with expression plasmids encoding the different peripherin isoforms together with plasmids encoding NF-L, NF-M, or NF-H. Assembly properties of the coexpressed proteins were analyzed by double immunofluorescence labeling using monoclonal antibody to peripherin (MAB1527; red) and polyclonal antiserum (green) to NF-L (AB1983), NF-M (AB1981), and NF-H (AB 1982) in the respective cotransfected cells. Isoforms Per 58 and Per 56 were capable of coassembly with all three neurofilament subunits (A, A′; B, B′; D, D′; E, E′; I, I′; and J, J′), whereas isoform Per 61 was incapable of coassembly into an IF network with any of the neurofilament subunits (C, C′; H, H′; and O, O′). Additional phenotypes were observed in cells cotransfected with Per 58 or Per 56 and NF-M or NF-H. These were filamentous bundles of peripherin in the presence of nonassembled NF-M (F, F′ and G, G′) or NF-H (K, K′ and L, L′; arrows in K indicate filaments) and nonassembled Per 58 or Per 56 in the presence of nonassembled NF-H (M, M′ and N, N′). Bar, 14 μm.

    Article Snippet: Transient transfections A human adrenal carcinoma cell line, SW13 vim(−), that lacks cytoplasmic IFs, was derived by dilutional cloning of SW13 cells obtained from the American Type Culture Collection.

    Techniques: Transfection, Expressing, Immunofluorescence, Labeling

    HDAC inhibitor treatment promotes paclitaxel resistance in SW13 cells. SW13 cells were left untreated (Control), or were treated with 1 nM FK228 for 24 h (FK228). After, 24 h, FK228 was removed, and control SW13 cells and FK228 pre-treated cells were plated into 96 well plates and left untreated (Control), or were treated with 1 nM, 10 nM, or 50 nM paclitaxel for ( a ) 24 h or ( b ) 48 h. Cell viability was assessed using an MTT assay. Data are presented as means ± SEM, and are representative of three independent experiments. Superscripts denote statistical significance, p

    Journal: BMC Cancer

    Article Title: Alterations in the glycome after HDAC inhibition impact oncogenic potential in epigenetically plastic SW13 cells

    doi: 10.1186/s12885-018-5129-4

    Figure Lengend Snippet: HDAC inhibitor treatment promotes paclitaxel resistance in SW13 cells. SW13 cells were left untreated (Control), or were treated with 1 nM FK228 for 24 h (FK228). After, 24 h, FK228 was removed, and control SW13 cells and FK228 pre-treated cells were plated into 96 well plates and left untreated (Control), or were treated with 1 nM, 10 nM, or 50 nM paclitaxel for ( a ) 24 h or ( b ) 48 h. Cell viability was assessed using an MTT assay. Data are presented as means ± SEM, and are representative of three independent experiments. Superscripts denote statistical significance, p

    Article Snippet: SW13 cell culture and treatment SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10% fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: MTT Assay

    HDAC inhibition induces changes in SW13 morphology, proliferation, and MMP activity. (A) Treatment with 1 nM FK228 significantly impacts SW13 cell morphology and actin (green) organization. (B and C) FK228 treatment decreases proliferation by nearly 90% as determined by the lower number of EdU positive cells (green) per total number of cells indicated by DAPI staining (blue). (D and E) FK228 treated cells exhibit higher levels of MMP2/9 (collagenase) activity as measured by increased fluorescence after incubation with a fluorescein-quenched gelatin substrate (DQ gel; green). Scale bars represent 10 μm

    Journal: BMC Cancer

    Article Title: Alterations in the glycome after HDAC inhibition impact oncogenic potential in epigenetically plastic SW13 cells

    doi: 10.1186/s12885-018-5129-4

    Figure Lengend Snippet: HDAC inhibition induces changes in SW13 morphology, proliferation, and MMP activity. (A) Treatment with 1 nM FK228 significantly impacts SW13 cell morphology and actin (green) organization. (B and C) FK228 treatment decreases proliferation by nearly 90% as determined by the lower number of EdU positive cells (green) per total number of cells indicated by DAPI staining (blue). (D and E) FK228 treated cells exhibit higher levels of MMP2/9 (collagenase) activity as measured by increased fluorescence after incubation with a fluorescein-quenched gelatin substrate (DQ gel; green). Scale bars represent 10 μm

    Article Snippet: SW13 cell culture and treatment SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10% fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Inhibition, Activity Assay, Staining, Fluorescence, Incubation

    The expression of HSPGs and related growth factors are in HDAC inhibitor treated SW13 cells. Relative mRNA expression of glypican-3 (GPC3), serglycin (SRGN), granulin (GRN), and fibroblast growth factor receptor (FGFR) 1 and 2, in SW13 cells following treatment with 1 nM of the HDAC inhibitor FK228 for 24. *Denotes statistical difference from control, p

    Journal: BMC Cancer

    Article Title: Alterations in the glycome after HDAC inhibition impact oncogenic potential in epigenetically plastic SW13 cells

    doi: 10.1186/s12885-018-5129-4

    Figure Lengend Snippet: The expression of HSPGs and related growth factors are in HDAC inhibitor treated SW13 cells. Relative mRNA expression of glypican-3 (GPC3), serglycin (SRGN), granulin (GRN), and fibroblast growth factor receptor (FGFR) 1 and 2, in SW13 cells following treatment with 1 nM of the HDAC inhibitor FK228 for 24. *Denotes statistical difference from control, p

    Article Snippet: SW13 cell culture and treatment SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10% fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Expressing

    HDAC inhibition and sample preparation influence lectin binding in fixed cells. SW13 cells were left untreated (Control), or were treated with 1 nM FK228 for 24 h (FK228). Cells were then fixed and left unpermeabilized ( a ), or were permeabilized with 0.2% Triton-X ( b ) before incubation with 20 μg/ml FITC labeled lectins for ~ 2 h. To demonstrate specificity of lectin binding, wheat germ agglutinin (WGA) stained cells were incubated with WGA elution buffer (+ Comp) for 30 min. Permeabilization of fixed cells before incubation with FITC-labeled lectins can significantly impacts staining of some lectins. All samples were mounted with ProLong Gold anti-fade reagent with DAPI to stain nuclei and photographed using a Zeiss Axiovert apotome with a 40X objective lens and a uniform exposure at each wavelength

    Journal: BMC Cancer

    Article Title: Alterations in the glycome after HDAC inhibition impact oncogenic potential in epigenetically plastic SW13 cells

    doi: 10.1186/s12885-018-5129-4

    Figure Lengend Snippet: HDAC inhibition and sample preparation influence lectin binding in fixed cells. SW13 cells were left untreated (Control), or were treated with 1 nM FK228 for 24 h (FK228). Cells were then fixed and left unpermeabilized ( a ), or were permeabilized with 0.2% Triton-X ( b ) before incubation with 20 μg/ml FITC labeled lectins for ~ 2 h. To demonstrate specificity of lectin binding, wheat germ agglutinin (WGA) stained cells were incubated with WGA elution buffer (+ Comp) for 30 min. Permeabilization of fixed cells before incubation with FITC-labeled lectins can significantly impacts staining of some lectins. All samples were mounted with ProLong Gold anti-fade reagent with DAPI to stain nuclei and photographed using a Zeiss Axiovert apotome with a 40X objective lens and a uniform exposure at each wavelength

    Article Snippet: SW13 cell culture and treatment SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10% fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Inhibition, Sample Prep, Binding Assay, Incubation, Labeling, Whole Genome Amplification, Staining

    HDAC inhibitor treatment significantly impacts SW13 cell lectin binding and sulfation status. Total protein was isolated from SW13 control cells and SW13 cells that had been treated with 1 nM FK228 for 24 h ( n = 4 per group). a FK228 treated SW13 cells have significantly higher levels of GAG-chain sulfation compared to SW13 control cells. b Lysates were subjected to lectin microarray analysis. Histograms of the fluorescence intensities lectins which were differentially bound between SW13 control cells and FK228 treated cells are shown. Data are presented as means ± SEM. c Lectin blots were performed to confirm lectin array results of select lectins (ConA, LPA, GS-I, Lotus, AIA, MPA, and ECA) that were differentially bound. DBA and UAE-1 were stained as negative controls. *Denotes statistical difference compared to control SW13- cells, p

    Journal: BMC Cancer

    Article Title: Alterations in the glycome after HDAC inhibition impact oncogenic potential in epigenetically plastic SW13 cells

    doi: 10.1186/s12885-018-5129-4

    Figure Lengend Snippet: HDAC inhibitor treatment significantly impacts SW13 cell lectin binding and sulfation status. Total protein was isolated from SW13 control cells and SW13 cells that had been treated with 1 nM FK228 for 24 h ( n = 4 per group). a FK228 treated SW13 cells have significantly higher levels of GAG-chain sulfation compared to SW13 control cells. b Lysates were subjected to lectin microarray analysis. Histograms of the fluorescence intensities lectins which were differentially bound between SW13 control cells and FK228 treated cells are shown. Data are presented as means ± SEM. c Lectin blots were performed to confirm lectin array results of select lectins (ConA, LPA, GS-I, Lotus, AIA, MPA, and ECA) that were differentially bound. DBA and UAE-1 were stained as negative controls. *Denotes statistical difference compared to control SW13- cells, p

    Article Snippet: SW13 cell culture and treatment SW13 cells were obtained from American Type Culture Collection (ATCC; CCL-105) and maintained in high glucose DMEM supplemented with 10% fetal bovine serum and 10 U/ml penicillin/streptomycin at 37 °C in a humidity controlled incubator.

    Techniques: Binding Assay, Isolation, Microarray, Fluorescence, Staining

    HSD17B4 exerts tumor suppressive effect in ACC cells NCI-H295R and SW13 showing ( A ) constitutive HSD17B4 level; ( B ) robust knockdown (KD) of HSD17B4 using shRNAs in both cell lines and ( C ) robust upregulation of HSD17B4 in both cell lines; HSD17B4-KD and overexpression in NCI-H295 and SW13 cells effecting on ( D ) proliferation, ( E ) cell cycle and ( F ) cell apoptosis, respectively ( ** P

    Journal: Oncotarget

    Article Title: Overexpression of HSD17B4 exerts tumor suppressive function in adrenocortical carcinoma and is not associated with hormone excess

    doi: 10.18632/oncotarget.22827

    Figure Lengend Snippet: HSD17B4 exerts tumor suppressive effect in ACC cells NCI-H295R and SW13 showing ( A ) constitutive HSD17B4 level; ( B ) robust knockdown (KD) of HSD17B4 using shRNAs in both cell lines and ( C ) robust upregulation of HSD17B4 in both cell lines; HSD17B4-KD and overexpression in NCI-H295 and SW13 cells effecting on ( D ) proliferation, ( E ) cell cycle and ( F ) cell apoptosis, respectively ( ** P

    Article Snippet: Cell lines and culture The human adrenocortical cancer (NCI-H295R) and adrenocortical small cell carcinoma (SW13) cell lines were originally obtained from ATCC and cultured in DMEM medium (Thermo Scientific, Logan, UT, USA) supplemented with 20% fetal bovine serum.

    Techniques: Over Expression

    HSD17B4 overexpression is associated with p53 signaling alteration ( A ) reproduction of TCGA ACC dataset showing heatmap of p53 signaling related genes in cases with and without HSD17B4 overexpression; ( B ) MDM4 expression is significantly lower in cases with HSD17B4 overexpression; ( C ) ATR expression is significantly higher in cases with HSD17B4 overexpression; enriched genes were confirmed for protein level using western blotting in ( D ) H295R cells with and without HSD17B4-KD, and in ( E ) SW13 cells with and without HSD17B4 replenish ( * P

    Journal: Oncotarget

    Article Title: Overexpression of HSD17B4 exerts tumor suppressive function in adrenocortical carcinoma and is not associated with hormone excess

    doi: 10.18632/oncotarget.22827

    Figure Lengend Snippet: HSD17B4 overexpression is associated with p53 signaling alteration ( A ) reproduction of TCGA ACC dataset showing heatmap of p53 signaling related genes in cases with and without HSD17B4 overexpression; ( B ) MDM4 expression is significantly lower in cases with HSD17B4 overexpression; ( C ) ATR expression is significantly higher in cases with HSD17B4 overexpression; enriched genes were confirmed for protein level using western blotting in ( D ) H295R cells with and without HSD17B4-KD, and in ( E ) SW13 cells with and without HSD17B4 replenish ( * P

    Article Snippet: Cell lines and culture The human adrenocortical cancer (NCI-H295R) and adrenocortical small cell carcinoma (SW13) cell lines were originally obtained from ATCC and cultured in DMEM medium (Thermo Scientific, Logan, UT, USA) supplemented with 20% fetal bovine serum.

    Techniques: Over Expression, Expressing, Western Blot

    HSD17B4 altered cell motility in H295R but not in SW13 cells HSD17B4-KD and overexpression in NCI-H295 and SW13 cells effecting on ( A ) cell invasion; ( B ) cell migration; and ( C ) colony formation, respectively; ( D ) HSD17B4-KD induced increased in vivo tumor growth of NCI-H295R cells ( ** P

    Journal: Oncotarget

    Article Title: Overexpression of HSD17B4 exerts tumor suppressive function in adrenocortical carcinoma and is not associated with hormone excess

    doi: 10.18632/oncotarget.22827

    Figure Lengend Snippet: HSD17B4 altered cell motility in H295R but not in SW13 cells HSD17B4-KD and overexpression in NCI-H295 and SW13 cells effecting on ( A ) cell invasion; ( B ) cell migration; and ( C ) colony formation, respectively; ( D ) HSD17B4-KD induced increased in vivo tumor growth of NCI-H295R cells ( ** P

    Article Snippet: Cell lines and culture The human adrenocortical cancer (NCI-H295R) and adrenocortical small cell carcinoma (SW13) cell lines were originally obtained from ATCC and cultured in DMEM medium (Thermo Scientific, Logan, UT, USA) supplemented with 20% fetal bovine serum.

    Techniques: Over Expression, Migration, In Vivo

    Down-regulation of Daxx leads to up-regulation of Brg1-regulated CD44 and SCEL genes. SW13 cell line was transfected with siRNA (control, siDaxx #1 and siDaxx B+C+D, a combination of three distinct siRNAs) and then infected with recombinant adenoviruses (0.05, 0.1 or 0.15 μl) expressing either EGFP or EGFP-tagged Brg1. The cells were then subjected to both (a) Western blotting and (b) qRT-PCR analyses. The statistical significance was assessed by paired Student's t-test.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Multifunctional adaptor protein Daxx interacts with chromatin-remodelling ATPase Brg1

    doi: 10.1016/j.bbrep.2015.12.012

    Figure Lengend Snippet: Down-regulation of Daxx leads to up-regulation of Brg1-regulated CD44 and SCEL genes. SW13 cell line was transfected with siRNA (control, siDaxx #1 and siDaxx B+C+D, a combination of three distinct siRNAs) and then infected with recombinant adenoviruses (0.05, 0.1 or 0.15 μl) expressing either EGFP or EGFP-tagged Brg1. The cells were then subjected to both (a) Western blotting and (b) qRT-PCR analyses. The statistical significance was assessed by paired Student's t-test.

    Article Snippet: 2.1 Cell cultures SW13, HeLa and HEK293T cells (all ATCC) were cultivated in DMEM, and supplemented with 10% FBS and antibiotics (penicillin+streptomycin).

    Techniques: Transfection, Infection, Recombinant, Expressing, Western Blot, Quantitative RT-PCR

    Transient RARRES2 overexpression inhibits cell proliferation in HEK293 cells and leads to reduced cellular invasion in H295R and HEK293 cells. ( a ) Baseline RARRES2 and CMKLR1 gene expression levels were barely detectable in the cell lines tested using TaqMan qPCR. ( b ) The time course for RARRES2 expression after transient transfection in HEK293, H295R and SW13 cell lines. ( c ) in vitro cell proliferation curve for cell lines transiently transfected with a vector control or RARRES2. Transient overexpression of RARRES2 inhibited cell proliferation in HEK293 cells, but not in H295R or SW13 cells. Data were presented as mean ± SEM. *, P

    Journal: Oncogene

    Article Title: RARRES2 functions as a tumor suppressor by promoting β-catenin phosphorylation/degradation and inhibiting p38 phosphorylation in adrenocortical carcinoma

    doi: 10.1038/onc.2016.497

    Figure Lengend Snippet: Transient RARRES2 overexpression inhibits cell proliferation in HEK293 cells and leads to reduced cellular invasion in H295R and HEK293 cells. ( a ) Baseline RARRES2 and CMKLR1 gene expression levels were barely detectable in the cell lines tested using TaqMan qPCR. ( b ) The time course for RARRES2 expression after transient transfection in HEK293, H295R and SW13 cell lines. ( c ) in vitro cell proliferation curve for cell lines transiently transfected with a vector control or RARRES2. Transient overexpression of RARRES2 inhibited cell proliferation in HEK293 cells, but not in H295R or SW13 cells. Data were presented as mean ± SEM. *, P

    Article Snippet: Cell Lines, Transfection, and Recombinant Protein Treatment Human ACC cell lines H295R and SW13 were purchased from ATCC (Rockville, MD), cultured in DMEM supplemented with 1% insulin transferrin selenium and 2.5% NuSerum I (Corning, Bedford, MA), and authenticated by short-tandem repeat profiling.

    Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Transfection, In Vitro, Plasmid Preparation

    (A) Aldosterone production in human CAR47, SW13, and various NCI-H295-derived cell models including HAC cell lines. All cells were incubated for 48 h followed by aldosterone measurement in the experimental medium. Results represent a minimum of 3 independent

    Journal: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme

    Article Title: Comparison of Aldosterone Production among Human Adrenocortical Cell Lines

    doi: 10.1055/s-0031-1298019

    Figure Lengend Snippet: (A) Aldosterone production in human CAR47, SW13, and various NCI-H295-derived cell models including HAC cell lines. All cells were incubated for 48 h followed by aldosterone measurement in the experimental medium. Results represent a minimum of 3 independent

    Article Snippet: SW13 human adrenocortical cells were cultured in Leibovitz’s L-15 medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Grand Island, NY).

    Techniques: Derivative Assay, HAC Assay, Incubation

    Phase contrast photomicrographs of human CAR47, SW13, and various NCI-H295-derived cell models including HAC cell lines. Scale bar=100μm.

    Journal: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme

    Article Title: Comparison of Aldosterone Production among Human Adrenocortical Cell Lines

    doi: 10.1055/s-0031-1298019

    Figure Lengend Snippet: Phase contrast photomicrographs of human CAR47, SW13, and various NCI-H295-derived cell models including HAC cell lines. Scale bar=100μm.

    Article Snippet: SW13 human adrenocortical cells were cultured in Leibovitz’s L-15 medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Grand Island, NY).

    Techniques: Derivative Assay, HAC Assay

    Comparison of transcript levels for the enzymes needed for aldosterone biosynthesis (Panel A) and hormonal receptors (Panel B) in human CAR47, SW13, and the various NCI-H295-derived cell models including the HAC cell lines. RNA was isolated from each

    Journal: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme

    Article Title: Comparison of Aldosterone Production among Human Adrenocortical Cell Lines

    doi: 10.1055/s-0031-1298019

    Figure Lengend Snippet: Comparison of transcript levels for the enzymes needed for aldosterone biosynthesis (Panel A) and hormonal receptors (Panel B) in human CAR47, SW13, and the various NCI-H295-derived cell models including the HAC cell lines. RNA was isolated from each

    Article Snippet: SW13 human adrenocortical cells were cultured in Leibovitz’s L-15 medium (Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Grand Island, NY).

    Techniques: Derivative Assay, HAC Assay, Isolation

    Detection of BRG1-dependent Z-DNA formation in the human CSF1 promoter using the Z-probe. ( A ) SW13 cells were transfected with the Z-probe or Z mut -probe together with the BRG1 expression vector as indicated in the figure. Whole-cell lysates were separated using SDS–PAGE, and the protein expression was analyzed through immunoblotting using specific antibodies as indicated in the figure. ( B ) SW13 cells were transfected with BRG1 together with the Z-probe (lanes 1 and 2) or Z mut -probe (lane 3) expression vectors, and the CSF1 gene expression was analyzed using real-time PCR. The value of lane 1 was arbitrarily set as 1, and the relative expression levels were expressed as the means ± SEM of four independent assays. ( C ) A schematic figure of the human CSF1 gene locus and regions detected using ChIP analysis. +1: transcription start site; ZZZ: CSF1 -TG repeat region; Ex4: CSF1 Exon 4 region. ( D ) The SW13 cells were transfected with the Z-probe or Z mut -probe together with BRG1 expression vectors, and subsequently, a ChIP assay was performed using an anti-GFP antibody (black bars). Normal rabbit IgG was used as a negative control (gray bars). Fold Z-DNA formation was measured using real-time PCR with specific primer sets for the upstream region of the CSF1 promoter ( CSF1 -3600), the CSF1 -TG or CSF1 -Ex4 regions. The anti-GFP ChIP value in lane 1 is set as 1, and the relative binding values are expressed as the means ± SEM of four independent assays. # P

    Journal: Nucleic Acids Research

    Article Title: Nrf2 activation is associated with Z-DNA formation in the human HO-1 promoter

    doi: 10.1093/nar/gkt243

    Figure Lengend Snippet: Detection of BRG1-dependent Z-DNA formation in the human CSF1 promoter using the Z-probe. ( A ) SW13 cells were transfected with the Z-probe or Z mut -probe together with the BRG1 expression vector as indicated in the figure. Whole-cell lysates were separated using SDS–PAGE, and the protein expression was analyzed through immunoblotting using specific antibodies as indicated in the figure. ( B ) SW13 cells were transfected with BRG1 together with the Z-probe (lanes 1 and 2) or Z mut -probe (lane 3) expression vectors, and the CSF1 gene expression was analyzed using real-time PCR. The value of lane 1 was arbitrarily set as 1, and the relative expression levels were expressed as the means ± SEM of four independent assays. ( C ) A schematic figure of the human CSF1 gene locus and regions detected using ChIP analysis. +1: transcription start site; ZZZ: CSF1 -TG repeat region; Ex4: CSF1 Exon 4 region. ( D ) The SW13 cells were transfected with the Z-probe or Z mut -probe together with BRG1 expression vectors, and subsequently, a ChIP assay was performed using an anti-GFP antibody (black bars). Normal rabbit IgG was used as a negative control (gray bars). Fold Z-DNA formation was measured using real-time PCR with specific primer sets for the upstream region of the CSF1 promoter ( CSF1 -3600), the CSF1 -TG or CSF1 -Ex4 regions. The anti-GFP ChIP value in lane 1 is set as 1, and the relative binding values are expressed as the means ± SEM of four independent assays. # P

    Article Snippet: Cell culture Human adrenal carcinoma SW13, 293 T-REx-3xFLAG-human Nrf2/Z-probe* cells were maintained in Dulbecco’s modified Eagle medium (Sigma-Aldrich) containing 10% fetal bovine serum and 100 U/ml penicillin–streptomycin (Gibco).

    Techniques: Transfection, Expressing, Plasmid Preparation, SDS Page, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Negative Control, Binding Assay