Journal: PLoS ONE

Article Title: Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

doi: 10.1371/journal.pone.0070643

Figure Lengend Snippet: Live or isopropanol-inactivated dead cells of E. coli O157:H7 ATCC 700728 (5×10 7 CFU) were incubated in the dark with or without 40 µM PMA for 15, 30, or 60. Real-time PCR was performed using the lpfA or hlyA primer sets , . Each bar represents the mean ± stdev for three independent replicates.

Article Snippet: Therefore, primers were selected based on minimum Ct (a measure of sensitivity) for amplifying 1 ng of E. coli O157:H7 ATCC 700728 genomic DNA (the equivalent of 10 5 genomes) and maximum Ct (a measure of selectivity) for amplifying 1 ng of DNA isolated from the closely-related bacterial strain E. coli K12.

Techniques: Incubation, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

doi: 10.1371/journal.pone.0070643

Figure Lengend Snippet: Cell amounts were quantified by real-time PCR (total; black bars) or PMA real-time PCR (viable; open bars) in mixtures of 10 3 or 10 4 live cells mixed with 10 6 isopropanol-inactivated dead cells. Samples were treated with 40 µM PMA for 30 min. Estimates of E coli O157:H7 cell amounts were based on comparisons to standard curves constructed using E. coli O157:H7 ATCC 700728 genomic DNA. Each bar represents the mean ± stdev for three independent replicates.

Article Snippet: Therefore, primers were selected based on minimum Ct (a measure of sensitivity) for amplifying 1 ng of E. coli O157:H7 ATCC 700728 genomic DNA (the equivalent of 10 5 genomes) and maximum Ct (a measure of selectivity) for amplifying 1 ng of DNA isolated from the closely-related bacterial strain E. coli K12.

Techniques: Real-time Polymerase Chain Reaction, Construct

Journal: PLoS ONE

Article Title: Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

doi: 10.1371/journal.pone.0070643

Figure Lengend Snippet: E. coli O157:H7 cell amounts were measured by plate count (culturable), real-time PCR (total), and PMA real-time PCR (viable) methods. Plants were inoculated with a spray bottle (A) or by drop (B) and incubated at a relative humidity of 30%. The mean ± stdev of 15 lettuce plants or leaves at each sampling time for three replicate experiments is shown.

Article Snippet: Therefore, primers were selected based on minimum Ct (a measure of sensitivity) for amplifying 1 ng of E. coli O157:H7 ATCC 700728 genomic DNA (the equivalent of 10 5 genomes) and maximum Ct (a measure of selectivity) for amplifying 1 ng of DNA isolated from the closely-related bacterial strain E. coli K12.

Techniques: Real-time Polymerase Chain Reaction, Incubation, Sampling

Journal: PLoS ONE

Article Title: Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

doi: 10.1371/journal.pone.0070643

Figure Lengend Snippet: E. coli O157:H7 amounts were measured by plate count (culturable), real-time PCR (total), and PMA real-time PCR (viable). Viable amounts were below the detection limit (3.7 log CFU/plant or leaf) at 2 and 48 h. The mean ± stdev of 12 lettuce plants at each sampling time is shown.

Article Snippet: Therefore, primers were selected based on minimum Ct (a measure of sensitivity) for amplifying 1 ng of E. coli O157:H7 ATCC 700728 genomic DNA (the equivalent of 10 5 genomes) and maximum Ct (a measure of selectivity) for amplifying 1 ng of DNA isolated from the closely-related bacterial strain E. coli K12.

Techniques: Real-time Polymerase Chain Reaction, Sampling

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A, C,E) ChIP assays were carried out in HeLa cells stimulated with IFN-γ for 0–2 hrs. Cell lysates were immunoprecipitated (IP’d) with control antibody or with antibody to endogenous 19S ATPase S6a, Sug1, or S7 and associated DNA was isolated and analyzed by real-time PCR using primers and probe spanning the CIITApIV proximal promoter. Real time PCR values were normalized to the total amount of DNA in the reaction (Input). IP values are represented as ATPase binding to CIITApIV promoter DNA relative to unstimulated samples. (B,D,F) ChIP signal at the inactive gene CD4. The control IgG values were 0.004±0.001. Values for control IgG and either Sug1 IP, S7 IP or S6a IP represent the mean ± SEM of three biologically independent experiments * p<0.05.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Immunoprecipitation, Control, Isolation, Real-time Polymerase Chain Reaction, Binding Assay

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A–B, D–E) HeLa cells were stimulated with IFN-γ as indicated and were harvested four hrs post treatment with 10 µM MG132 or 10 µM Lactacystin. mRNA was extracted and cDNA was generated using indicated reverse primers followed by amplification via real-time PCR. CIITA mRNA transcripts were obtained using primers and probes specific for CIITA exon IV and exon VII were normalized to GAPDH. (C) 18S rRNA transcripts for control and MG132 treated cells were obtained using primers and probe specific for 18S rRNA and were normalized to GAPDH. The 18 hr control sample was set to 100%. Data shown represents the mean ± SEM of three biologically independent experiments.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Generated, Amplification, Real-time Polymerase Chain Reaction, Control

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: HeLa cells were transfected with HSE-Luciferase reporter, control siRNA, or S6a siRNA and were treated with MG132 six hrs prior to harvest. Cells were harvested following 48 hrs of incubation, lysed in cell lysis buffer, and analyzed by Luciferase assay. Luciferase readings obtained were normalized by Bradford assay. Data shown represents values obtained from three independent experiments. The negative control is a mixture of non-inducible reporter construct and constitutively expressing Renilla luciferase construct. The positive control is an inducible transcription factor-responsive construct expressing firefly luciferase, and a constitutively expressing Renilla luciferase construct.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Luciferase, Control, Incubation, Lysis, Bradford Assay, Negative Control, Construct, Expressing, Positive Control

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A–I) ChIP assays were carried out in HeLa cells stimulated with IFN-γ for 0–2 hrs. Cell lysates were IP’d with control antibody or with antibody to endogenous Sug1 (A and B), S7 (D and E), or S6a (G and H) and associated DNA was isolated and analyzed by real-time PCR using primers and probes spanning CIITApIV exon IV (A, C, E) and exon VII (B, D, F). Real time PCR IP values were normalized to the total amount of DNA (input); IP values are represented as ATPase binding to CIITApIV exon IV or exon VII DNA relative to unstimulated samples. (C,F,I) ChIP signal at the inactive gene CD4. The control IgG values were 0.005±0.001. Values for control and IP represent mean ± SEM of three biologically independent experiments. *p<0.05, **p<0.005. G. Mobility shift assay of Sug1 with a 90 nucleotide single stranded DNA on a native 8% polyacrylamide gel with a tris-borate magnesium running buffer; 0.7 µM DNA, 0.85 µM sug1, and 500 µM ATP. DNA was visualized with SYBER Green II dye.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Control, Isolation, Real-time Polymerase Chain Reaction, Binding Assay, Mobility Shift

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A–C) HeLa cells were co-transfected with Myc tagged S6a, S7, or Sug1 and Flag tagged Hexim or HA tagged CDK9 as indicated. Cells were lysed and IP’d with Myc beads (lane 1) as a positive control, mouse isotype IgG (lane 2) as a negative control, flag beads (lane 3), and HA beads (lane 4). IP samples (top panel) and lysates (bottom panel) were IB’d for Myc, Flag, and HA as indicated. (D–E) HeLa cells were lysed and IP’d with either Hexim or CDK9 (lane 1) as a positive control, mouse isotype IgG (lane 2) as a negative control, or with S6a (lane 3), S7 (lane 4), and Sug1 (lane 5). IP samples (top panel) and lysates (bottom panel) were IB’d for Hexim or CDK9 as indicated. Results shown are indicative of data from three biologically independent experiments.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A) HeLa cells were transfected with myc tagged S6a, S7 or Sug1 as indicated. Cells were lysed and IP’d with myc beads (first lane, top panels) as a positive control, with mouse isotype IgG (second lane, top panels) as a negative control, and with Ser5p-RNA Pol II antibody (third lane, top panels). IP’d samples (top panels) and lysates (middle and bottom panels) were IB for myc ATPases or for Ser5p-RNA pol II as indicated. (B) HeLa cells were lysed and IP’d with Ser5p-RNA Pol II (lane 1) as a positive control, mouse isotype IgG (lane 2) as a negative control, or with S6a (lane 3), S7 (lane 4), or Sug1 (lane 5). IP samples (top panel) and lysates (bottom panel) were IB’d Ser5p-RNA Pol II as indicated. Results shown are indicative of data from three biologically independent experiments.

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Positive Control, Negative Control

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: A. HeLa cells were transfected with siRNA or were treated with proteasome inhibitors as indicated. Cells were harvested following 48hrs of siRNA incubation. Cell lysates were IB’d with Ser2p-RNA pol II antibody (top panels), Ser5p-RNA pol II (middle panels), or with RNA pol II antibody (bottom panels). Cells treated with proteasome inhibitors serve as a positive control for degradation dependent effects. Results shown are indicative of data from three biologically independent experiments. Sug1, S7, and S6a protein expression was effectively decreased using specific siRNA. Actin blots demonstrate loading and siRNA specificity controls .

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Incubation, Positive Control, Expressing

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A,C,E) ChIP assays were carried out in HeLa cells transfected with ATPase specific or with control siRNA and stimulated with IFN-γ for 0–2 hrs. Cell lysates were crosslinked, sonicated, lysed, and IP’d with either antibody against endogenous RNA pol II or with control antibody (IgG). Associated DNA was analyzed via real-time PCR using primers and probe specific for the CIITApIV proximal promoter. Real time PCR IP values were normalized to total amount of reaction DNA (Input). The values for control IP and RNA Pol II IP represent the mean of three biologically independent experiments *p<0.05, **p<0.005, ***p<0.0005 versus control siRNA. (B, D, F) ChIP signal at the inactive gene CD4. Sug1, S7, and S6a protein expression was effectively decreased using ATPase specific siRNA .

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Control, Sonication, Real-time Polymerase Chain Reaction, Expressing

Journal: PLoS ONE

Article Title: Nonproteolytic Roles of 19S ATPases in Transcription of CIITApIV Genes

doi: 10.1371/journal.pone.0091200

Figure Lengend Snippet: (A, C, E) ChIP assays were carried out in HeLa cells transfected with ATPase specific or with control siRNA and stimulated with IFN-γ for 0–2 hrs. Cell lysates were crosslinked, sonicated, lysed, and IP’d with either antibody against endogenous TBP or with control antibody (IgG). Associated DNA was analyzed via real-time PCR using primers and probe specific for the CIITApIV proximal promoter. Real time PCR IP values were normalized to total amount of reaction DNA (Input). The values for control IP and TBP IP are representative data *p<0.05, **p<0.005, ***p<0.0005 versus control siRNA. (B, D, F) ChIP signal at the inactive gene CD4. Sug1, S7, and S6a protein expression was effectively decreased using ATPase specific siRNA .

Article Snippet: HeLa cells (human epithelial) from ATCC (Manassas, VA) were cultured using high-glucose Dulbecco modified Eagle (DMEM) medium (Mediatech Inc., Herndon, VA) supplemented with 10% fetal bovine serum, 5 mM penicillin-streptomycin, and 5 mM L-glutamine.

Techniques: Transfection, Control, Sonication, Real-time Polymerase Chain Reaction, Expressing

Journal: Cell & Bioscience

Article Title: The role of GLI1 for 5-Fu resistance in colorectal cancer

doi: 10.1186/s13578-017-0145-7

Figure Lengend Snippet: Characterization of 5-FU resistant LoVo-R cells. a Shows morphology of LoVo and LoVo-R cells. b Shows the IC50 dose of 5-FU calculated from measurement of cell viability in different concentrations of 5-FU (48 h). The X-axis is 5-FU concentration (μg/ml), and the Y-axis is O.D. values. Significant difference was indicated by *p < 0.05, **p < 0.005, or ***p < 0.0005

Article Snippet: The human colorectal cancer cell line LoVo was purchased from ATCC.

Techniques: Concentration Assay

Journal: Cell & Bioscience

Article Title: The role of GLI1 for 5-Fu resistance in colorectal cancer

doi: 10.1186/s13578-017-0145-7

Figure Lengend Snippet: Up-regulated of GLI1 signaling axis in LoVo-R (in comparison with LoVo) cells. After next generation sequencing, we performed ingenuity pathway analysis (IPA). a Shows up-regulation of GLI1 and its signaling molecules, and the up-regulated genes are in red . b Detection of GLI1 protein in LoVo and LoVo-R cells. β-actin was used as the internal control

Article Snippet: The human colorectal cancer cell line LoVo was purchased from ATCC.

Techniques: Comparison, Next-Generation Sequencing, Control

Journal: Cell & Bioscience

Article Title: The role of GLI1 for 5-Fu resistance in colorectal cancer

doi: 10.1186/s13578-017-0145-7

Figure Lengend Snippet: The effect of GLI1/2 knockdown on 5-FU response in LoVo-R cells. a Real-time PCR detection of GLI1 after GLI1 shRNA expression. b Detection of GLI1 protein by Western blotting. c The effect of GLI1 -ShRNAs (shown as shGLI1) on the IC50 of 5-FU (measured as shown in Fig. b). d Effects of GLI2 shRNAs (shown as shGLI2) on 5-FU response. Significant difference was indicated by *p < 0.05, **p < 0.005, or ***p < 0.0005

Article Snippet: The human colorectal cancer cell line LoVo was purchased from ATCC.

Techniques: Knockdown, Real-time Polymerase Chain Reaction, shRNA, Expressing, Western Blot

Journal: Cell & Bioscience

Article Title: The role of GLI1 for 5-Fu resistance in colorectal cancer

doi: 10.1186/s13578-017-0145-7

Figure Lengend Snippet: The effects of GLI1 knockdown on gene/protein expression. a Real-time analysis of GLI1, GLI2, Snai1 and Snai2 in GLI2 shRNAs-expressing LoVo-R cells. Significant difference was indicated by ***p < 0.0005. b Effects of Gli2 shRNAs on vimentin expression. c The effect of GLI1 knockdown on Sox2 and CD44 proteins

Article Snippet: The human colorectal cancer cell line LoVo was purchased from ATCC.

Techniques: Knockdown, Expressing

Journal: Cell & Bioscience

Article Title: The role of GLI1 for 5-Fu resistance in colorectal cancer

doi: 10.1186/s13578-017-0145-7

Figure Lengend Snippet: Cell invasiveness assay. Different cells were subjected to cell invasiveness as described in the methods, and invasive cells were visualized by blue in the staining ( a ). b Shows the summary from three independent experiments, with the value (number of invasive cells) from LoVo cells as 100

Article Snippet: The human colorectal cancer cell line LoVo was purchased from ATCC.

Techniques: Staining

Journal: Oncology Letters

Article Title: miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1

doi: 10.3892/ol.2017.5709

Figure Lengend Snippet: miR-150 is downregulated in OS patient tissues and cell lines. (A) Relative expression levels of miR-150 in 40 OS patient tissue samples and adjacent normal tissue sample counterparts. The relative expression levels of miR-150 were determined using RT-qPCR analysis. (B) Relative expression levels of miR-150, determined using RT-qPCR analysis in U2OS, MG63 and SaOS2 OS cell lines and the normal hFOB1.19 human OS cell line. Student's t -test was used to analyze significant differences.*P<0.05. Data are presented as the mean + standard deviation. OS, osteosarcoma; miR, microRNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: The SaOS2, U2OS and MG63 human OS cell lines and the hFOB1.19 normal human osteoblast cell line were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Standard Deviation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1

doi: 10.3892/ol.2017.5709

Figure Lengend Snippet: miR-150 overexpression is inversely correlated with OS cell growth. (A) U2OS, (B) MG63 and (C) SaOS2 OS cell lines were transfected with miR-150 and the numbers of cells were determined at different time points using a fluorescence-based CyQUANT cell proliferation assay kit. In all cases, Student's t -test was used to analyze significant differences, *P<0.05. Data are presented as the mean + standard deviation. OS, osteosarcoma; miR, microRNA; NC, negative control.

Article Snippet: The SaOS2, U2OS and MG63 human OS cell lines and the hFOB1.19 normal human osteoblast cell line were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Over Expression, Transfection, Fluorescence, CyQUANT Assay, Proliferation Assay, Standard Deviation, Negative Control

Journal: Oncology Letters

Article Title: miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1

doi: 10.3892/ol.2017.5709

Figure Lengend Snippet: miR-150 overexpression is inversely correlated with OS cell migration and invasion. Transwell migration and invasion assays of (A) U2OS, (B) MG63 and (C) SaOS2 OS cell lines. The assays were performed following transfection of the cell lines with miR-150. Representative images of migrated and invaded cells on the membrane (magnification, ×100). OS, osteosarcoma; miR, microRNA; NC, negative control.

Article Snippet: The SaOS2, U2OS and MG63 human OS cell lines and the hFOB1.19 normal human osteoblast cell line were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Over Expression, Migration, Transfection, Membrane, Negative Control

Journal: Oncology Letters

Article Title: miR-150 is downregulated in osteosarcoma and suppresses cell proliferation, migration and invasion by targeting ROCK1

doi: 10.3892/ol.2017.5709

Figure Lengend Snippet: miR-150 negatively regulates gene expression of ROCK1 in OS cell lines. (A) Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression of ROCK1 in U2OS, MG63 and SaOS2 OS cell lines overexpressing miR150. Student's t -test was used to analyze significant differences. *P<0.05. Data are presented as the mean + standard deviation. (B) Western blot analysis of protein expression of ROCK1 in U2OS, MG63 and SaOS2 OS cell lines overexpressing miR150. A corresponding NC was used in the absence of miR-150. β-actin protein was used as an additional control. miR, microRNA; ROCK1, Rho-associated kinase 1; NC, negative control.

Article Snippet: The SaOS2, U2OS and MG63 human OS cell lines and the hFOB1.19 normal human osteoblast cell line were obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation, Western Blot, Control, Negative Control

Journal: FEBS Open Bio

Article Title: Doxorubicin promotes breast cancer cell migration and invasion via DCAF13

doi: 10.1002/2211-5463.13330

Figure Lengend Snippet: DCAF13 promotes EMT in human breast cancer. (A) Hierarchical cluster analysis of differentially expressed genes (|logFC| > 1 and P adj < 0.001) between DCAF13_high and DCAF13_low breast cancer samples. (B) The volcano plot of differential gene expression between DCAF13_high and DCAF13_low breast cancer samples. The horizontal dashed line marks the threshold ( P adj < 0.05) for defining a gene as significantly upregulated or downregulated. The vertical dashed lines represent two‐fold differences in expression. Significantly differentially expressed genes are shown as red dots ( P adj < 0.05 and |log2FC| > 1). (C) GSEA analysis was performed to identify the pathways altered in DCAF13_high samples compared to DCAF13_low samples. (D) A quantitative PCR was carried out to verify DCAF13 knockdown efficiency in BT549 cells and MDA‐MB‐231 cells. Data are presented as the mean ± SD. ( n = 3 independent experiments). *** P < 0.001, unpaired t ‐test. (E,F) Western blot and immunofluorescence staining assays were carried out to examine the effect of DCAF13 knockdown on EMT marker gene expression. Scale bar = 50 μm.

Article Snippet: Breast cancer cell line BT549 and MDA‐MB‐231 were originally purchased from the ATCC (Manassas, VA, USA).

Techniques: Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Knockdown, Western Blot, Immunofluorescence, Staining, Marker

Journal: FEBS Open Bio

Article Title: Doxorubicin promotes breast cancer cell migration and invasion via DCAF13

doi: 10.1002/2211-5463.13330

Figure Lengend Snippet: Doxorubicin promotes breast cancer cell migration, invasion and EMT via DCAF13. (A) Doxorubicin treatment increased DCAF13 expression in breast cancer cells. BT549 cells and MDA‐MB‐231 cells were treated with doxorubicin and total RNA was extracted for real‐time PCR analysis. Three independent experiments were carried out and the results were quantified and represented as the mean ± SD. *** P < 0.001, unpaired t ‐test. (B,C) Doxorubicin promotes breast cancer cell migration via DCAF13. Wound healing assays were carried out to assess the effect of doxorubicin treatment on breast cancer cell migration and the involvement of DCAF13. Three independent experiments were carried out and the results were quantified and represented as the mean ± SD. ** P < 0.01, unpaired t ‐test. Scale bar = 100 μm. (D,E) Doxorubicin promotes breast cancer cell invasion via DCAF13. Transwell cell invasion assays were performed to assess the effect of doxorubicin treatment on breast cancer cell invasion and the involvement of DCAF13. Three independent experiments were carried out and the results were quantified and represented as the mean ± SD. ** P < 0.01, *** P < 0.001, unpaired t ‐test. Scale bar = 100 μm. (F) Doxorubicin promotes the EMT in breast cancer cells via DCAF13. A western blot assay was performed to examine the effect of doxorubicin treatment on EMT marker gene expression and the involvement of DCAF13.

Article Snippet: Breast cancer cell line BT549 and MDA‐MB‐231 were originally purchased from the ATCC (Manassas, VA, USA).

Techniques: Migration, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Marker, Gene Expression