anti sox2  (Thermo Fisher)


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    Name:
    SOX2 Monoclonal Antibody 20G5
    Description:
    SOX2 Monoclonal Antibody for Western Blot IF ICC IHC P Flow IP ChIP
    Catalog Number:
    ma1014
    Price:
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    Category:
    Antibodies Secondary Detection Reagents
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    Structured Review

    Thermo Fisher anti sox2
    Immunohistochemistry of lung atypical carcinoid. <t>Anti-SOX2</t> monoclonal antibody of tumor slides. Representative images were shown in 20× and 40× magnification.
    SOX2 Monoclonal Antibody for Western Blot IF ICC IHC P Flow IP ChIP
    https://www.bioz.com/result/anti sox2/product/Thermo Fisher
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    anti sox2 - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Cancer stem-neuroendocrine cells in an atypical carcinoid case report"

    Article Title: Cancer stem-neuroendocrine cells in an atypical carcinoid case report

    Journal: Translational Lung Cancer Research

    doi: 10.21037/tlcr.2019.12.07

    Immunohistochemistry of lung atypical carcinoid. Anti-SOX2 monoclonal antibody of tumor slides. Representative images were shown in 20× and 40× magnification.
    Figure Legend Snippet: Immunohistochemistry of lung atypical carcinoid. Anti-SOX2 monoclonal antibody of tumor slides. Representative images were shown in 20× and 40× magnification.

    Techniques Used: Immunohistochemistry

    2) Product Images from "Embryonic Cerebrospinal Fluid Increases Neurogenic Activity in the Brain Ventricular-Subventricular Zone of Adult Mice"

    Article Title: Embryonic Cerebrospinal Fluid Increases Neurogenic Activity in the Brain Ventricular-Subventricular Zone of Adult Mice

    Journal: Frontiers in Neuroanatomy

    doi: 10.3389/fnana.2017.00124

    Effect of Embryonic CSF (E-CSF) on mitotic activity in SVZ niche NSCs, monitored by nuclear incorporation of BrdU (red). Confocal photomicrographs ( A,B show the SVZ of “ in vitro ” cultured adult mice brain slices (see Figure 1B ). Note the substantial increase in NSCs mitotic activity under the ventricular surface (white arrows) induced by E-CSF (B) , compared with the controls (A) . Confocal photomicrographs (C–E) show the Striatum area of “ in vitro ” cultured adult mice brain slices close to the SVZ (ST in Figure 1B ). Quantification of BrdU positive nuclei in Control (C) , E-CSF (D) and A-CSF (E) treated brain slices was plotted in graph bars (F) and expressed as means ± SD ( n = 20); the significant threshold was set at p ≤ 0.001 (*) according to the one-way ANOVA, post hoc Bonferroni test. The results reveal a statistically significant increase (49%) in the number of BrdU positive NSCs in CSF-E treated brain slices with respect to the controls and A-CSF treated slices; this suggests a specific activation by E-CSF of mitotic activity in NSCs of the SVZ niche. Confocal photomicrographs (G,I) correspond to the striatum area of “ in vitro ” cultured adult mice brain slices, close to SVZ (ST in Figure 1B ). Images show double immunolabeling with antiBrdU (green) and antiSox2 (red) antibody; co-localization of both antibodies was interpreted as NSCs dividing (BrdU positive) but not differentiating (Sox2 positive). Quantification of co-labeled BrdU and Sox2 NSCs in Control (G) and E-CSF treated brain slices (I) was plotted in graph bars (H) and expressed as means ± SD ( n = 20); the significant threshold was set at p ≤ 0.001 (*) according to the two-tailed Student’s t -test. The results reveal a statistically significant increase (141%) in the number of NSCs that undergo replication but remain undifferentiated in CSF-E treated brain slices, in comparison with the controls. Scale bar: 25 μm (A–E) and 10 μm (G,I) .
    Figure Legend Snippet: Effect of Embryonic CSF (E-CSF) on mitotic activity in SVZ niche NSCs, monitored by nuclear incorporation of BrdU (red). Confocal photomicrographs ( A,B show the SVZ of “ in vitro ” cultured adult mice brain slices (see Figure 1B ). Note the substantial increase in NSCs mitotic activity under the ventricular surface (white arrows) induced by E-CSF (B) , compared with the controls (A) . Confocal photomicrographs (C–E) show the Striatum area of “ in vitro ” cultured adult mice brain slices close to the SVZ (ST in Figure 1B ). Quantification of BrdU positive nuclei in Control (C) , E-CSF (D) and A-CSF (E) treated brain slices was plotted in graph bars (F) and expressed as means ± SD ( n = 20); the significant threshold was set at p ≤ 0.001 (*) according to the one-way ANOVA, post hoc Bonferroni test. The results reveal a statistically significant increase (49%) in the number of BrdU positive NSCs in CSF-E treated brain slices with respect to the controls and A-CSF treated slices; this suggests a specific activation by E-CSF of mitotic activity in NSCs of the SVZ niche. Confocal photomicrographs (G,I) correspond to the striatum area of “ in vitro ” cultured adult mice brain slices, close to SVZ (ST in Figure 1B ). Images show double immunolabeling with antiBrdU (green) and antiSox2 (red) antibody; co-localization of both antibodies was interpreted as NSCs dividing (BrdU positive) but not differentiating (Sox2 positive). Quantification of co-labeled BrdU and Sox2 NSCs in Control (G) and E-CSF treated brain slices (I) was plotted in graph bars (H) and expressed as means ± SD ( n = 20); the significant threshold was set at p ≤ 0.001 (*) according to the two-tailed Student’s t -test. The results reveal a statistically significant increase (141%) in the number of NSCs that undergo replication but remain undifferentiated in CSF-E treated brain slices, in comparison with the controls. Scale bar: 25 μm (A–E) and 10 μm (G,I) .

    Techniques Used: Activity Assay, In Vitro, Cell Culture, Mouse Assay, Activation Assay, Immunolabeling, Labeling, Two Tailed Test

    3) Product Images from "Six1 is essential for differentiation and patterning of the mammalian auditory sensory epithelium"

    Article Title: Six1 is essential for differentiation and patterning of the mammalian auditory sensory epithelium

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1006967

    Temporal deletion of Six1 between E12.75–13.5 blocks hair cell induction. (A) Whole-mount Myo7a staining of E18.5 cochleae showing four rows of hair cells along the entire cochlea in Eya1 CreER mice but extra inner hair cells present from middle toward apex (arrows). In Six1 CKO cochlea, the organ of Corti appears narrower and hair cells show abnormal morphology with gradually decreased outer rows of hair cells from four in the base and one row in the apex. (B) Co-immunostaining for Myo7a and Sox2 on sections from P0 cochleae treated with tamoxifen between E13.5–14.5 showing one inner and three outer hair cells in the base, one inner and two outer hair cells in the middle and only one hair cells in the apex in Six1 CKO. All Myo7a + hair cells are also Sox2 + in the CKO mutant. Scale bars: 30 μm.
    Figure Legend Snippet: Temporal deletion of Six1 between E12.75–13.5 blocks hair cell induction. (A) Whole-mount Myo7a staining of E18.5 cochleae showing four rows of hair cells along the entire cochlea in Eya1 CreER mice but extra inner hair cells present from middle toward apex (arrows). In Six1 CKO cochlea, the organ of Corti appears narrower and hair cells show abnormal morphology with gradually decreased outer rows of hair cells from four in the base and one row in the apex. (B) Co-immunostaining for Myo7a and Sox2 on sections from P0 cochleae treated with tamoxifen between E13.5–14.5 showing one inner and three outer hair cells in the base, one inner and two outer hair cells in the middle and only one hair cells in the apex in Six1 CKO. All Myo7a + hair cells are also Sox2 + in the CKO mutant. Scale bars: 30 μm.

    Techniques Used: Staining, Mouse Assay, Immunostaining, Mutagenesis

    Deletion of Six1 in the developing cochlea using Eya1 CreER or Sox2 CreER leads to shortened and thickened prosensory primordium. Cochleae were dissected from E14.5 embryos given tamoxifen at E11.5 (9 am) and E12.5 (9 am) and processed for whole-mount (A-D) or section (E-G) immunostaining with anti-Sox2 (green) and -p27 Kip1 (red). Hoechst was used for nuclear-counter staining. (E-G) Section collected from mid-cochlear duct in wild-type, Eya1 CreER or Sox2 CreER ;Six1 fl/fl ( Cko/Cko ) littermates as indicated by dashed line in A, B, D respectively. Bracket indicates p27 Kip1 -positive prosensory domain within the cochlea epithelium and its width on mediolateral axis is comparable between control and mutant littermates. (H) Spatial calibration of Sox2 + and p27 Kip1+ width, height and square area and value represents average number (±standard deviations) per section (6 μm) (see Methods for calibration). P- value was measured for +/+ and Cko/Cko using Two-tailed Student’s t-test. Scale bars: 200 μm in A-C and 40 μm in D,E.
    Figure Legend Snippet: Deletion of Six1 in the developing cochlea using Eya1 CreER or Sox2 CreER leads to shortened and thickened prosensory primordium. Cochleae were dissected from E14.5 embryos given tamoxifen at E11.5 (9 am) and E12.5 (9 am) and processed for whole-mount (A-D) or section (E-G) immunostaining with anti-Sox2 (green) and -p27 Kip1 (red). Hoechst was used for nuclear-counter staining. (E-G) Section collected from mid-cochlear duct in wild-type, Eya1 CreER or Sox2 CreER ;Six1 fl/fl ( Cko/Cko ) littermates as indicated by dashed line in A, B, D respectively. Bracket indicates p27 Kip1 -positive prosensory domain within the cochlea epithelium and its width on mediolateral axis is comparable between control and mutant littermates. (H) Spatial calibration of Sox2 + and p27 Kip1+ width, height and square area and value represents average number (±standard deviations) per section (6 μm) (see Methods for calibration). P- value was measured for +/+ and Cko/Cko using Two-tailed Student’s t-test. Scale bars: 200 μm in A-C and 40 μm in D,E.

    Techniques Used: Immunostaining, Staining, Mutagenesis, Two Tailed Test

    Model illustrating the role of Six1 in regulating auditory sensory cell development. This study demonstrates that Six1 regulates proliferation of sensory progenitors in the cochlea epithelium. While a direct interaction between Eya1/Six1/Sox2 proteins coordinately regulates Atoh1 expression in cochlear explant, this study provides in vivo evidence supporting a role for Six1 in hair cell fate specification and Atoh1 expression. In differentiating hair cells, our data show that Six1 activity is necessary for downregulation of Sox2 and maintenance of Fgf8 expression.
    Figure Legend Snippet: Model illustrating the role of Six1 in regulating auditory sensory cell development. This study demonstrates that Six1 regulates proliferation of sensory progenitors in the cochlea epithelium. While a direct interaction between Eya1/Six1/Sox2 proteins coordinately regulates Atoh1 expression in cochlear explant, this study provides in vivo evidence supporting a role for Six1 in hair cell fate specification and Atoh1 expression. In differentiating hair cells, our data show that Six1 activity is necessary for downregulation of Sox2 and maintenance of Fgf8 expression.

    Techniques Used: Expressing, In Vivo, Activity Assay

    Altered cell proliferation in Six1 CKO ( Eya1 CreER ) cochlea epithelium. (A) Immunostaining for Sox2 (green) and EdU (red) on sections of basal and middle regions of cochleae from E17.5 embryos given EdU at E14.5 and tamoxifen at E11.5–12.5. Arrows point to high levels of Sox2 in cells within the lumenal layer. (B) Immunostaining for Sox2 (green) and EdU (red) on sections of middle region of cochleae from E14.5 embryos given tamoxifen at E11.5–12.5 and EdU at E11.5. (C) Number of EdU + Sox2 + cells per section (6 μm) or EdU + cells per cochlea (see Methods for quantification). P -value was measured for +/+ and Cko/Cko using Two-tailed Student’s t-test: p
    Figure Legend Snippet: Altered cell proliferation in Six1 CKO ( Eya1 CreER ) cochlea epithelium. (A) Immunostaining for Sox2 (green) and EdU (red) on sections of basal and middle regions of cochleae from E17.5 embryos given EdU at E14.5 and tamoxifen at E11.5–12.5. Arrows point to high levels of Sox2 in cells within the lumenal layer. (B) Immunostaining for Sox2 (green) and EdU (red) on sections of middle region of cochleae from E14.5 embryos given tamoxifen at E11.5–12.5 and EdU at E11.5. (C) Number of EdU + Sox2 + cells per section (6 μm) or EdU + cells per cochlea (see Methods for quantification). P -value was measured for +/+ and Cko/Cko using Two-tailed Student’s t-test: p

    Techniques Used: Immunostaining, Two Tailed Test

    Downregulation of Sox2 in differentiating hair cells is disrupted in Six1 CKO. (A) Immunostaining for Sox2 (green) on whole-mount and (B) Sox2 (green)/Myo7a (red) on sections of cochleae from wild-type or Six1 CKO ( Eya1 CreER ;Six1 fl/fl ) littermate embryos at E18.5 (given tamoxifen at E11.5–12.5). Cochlear section region was indicated by dashed line in A. Arrows point to Sox2 expression in the GER cells flanking the inner hair cells. (C) Atoh1 in situ hybridization showing Atoh1 expression in the organ of Corti in wild-type embryos at E17.5 and absence of Atoh1 in Atoh1 fl/fl ;Eya1 CreER littermate embryos (arrow) given tamoxifen at E14.75–15.5. (D) Immunostaining for Myo7a (green) and Sox2 (red) on cochlear sections from E17.5 wild-type or Eya1 CreER ;Atoh1 fl/fl littermate embryos given tamoxifen at E14.75–15.5. (E) Immunostaining for Pou4f3 (green) and Six1 (red) on cochlear sections from E17.5 wild-type and Eya1 CreER ;Atoh1 fl/fl littermate embryos given tamoxifen at E14.75–15.5. Scale bars: 200 μm in A; 30 μm in B,D,E; 40 μm in C.
    Figure Legend Snippet: Downregulation of Sox2 in differentiating hair cells is disrupted in Six1 CKO. (A) Immunostaining for Sox2 (green) on whole-mount and (B) Sox2 (green)/Myo7a (red) on sections of cochleae from wild-type or Six1 CKO ( Eya1 CreER ;Six1 fl/fl ) littermate embryos at E18.5 (given tamoxifen at E11.5–12.5). Cochlear section region was indicated by dashed line in A. Arrows point to Sox2 expression in the GER cells flanking the inner hair cells. (C) Atoh1 in situ hybridization showing Atoh1 expression in the organ of Corti in wild-type embryos at E17.5 and absence of Atoh1 in Atoh1 fl/fl ;Eya1 CreER littermate embryos (arrow) given tamoxifen at E14.75–15.5. (D) Immunostaining for Myo7a (green) and Sox2 (red) on cochlear sections from E17.5 wild-type or Eya1 CreER ;Atoh1 fl/fl littermate embryos given tamoxifen at E14.75–15.5. (E) Immunostaining for Pou4f3 (green) and Six1 (red) on cochlear sections from E17.5 wild-type and Eya1 CreER ;Atoh1 fl/fl littermate embryos given tamoxifen at E14.75–15.5. Scale bars: 200 μm in A; 30 μm in B,D,E; 40 μm in C.

    Techniques Used: Immunostaining, Expressing, In Situ Hybridization

    4) Product Images from "Dynamic regulation of chromatin accessibility by pluripotency transcription factors across the cell cycle"

    Article Title: Dynamic regulation of chromatin accessibility by pluripotency transcription factors across the cell cycle

    Journal: eLife

    doi: 10.7554/eLife.50087

    Correlation between OCT4 binding and chromatin accessiblity, and analysis of results from random forest model. ( A ) Correlation between the log of normalized OCT4 ChIP-seq reads per bp (x-axis) and the log2 fold-change values of accessibility loss upon OCT4 depletion (y-axis) at all OCT4 binding sites in ZHBTc4 cells. ( B ) Correlation between the log of normalized OCT4 ChIP-seq reads per bp (x-axis) and the log of normalized ATAC-seq reads per bp (y-axis) at all OCT4 binding sites in ZHBTc4 cells. Coefficient (R) and p-values are based on the Pearson correlation coefficient. ( C ) Cluster predictions of regions in the test data based on a random forest model using mouse ES cell ChIP-seq data (see Materials and methods). The x‐axis shows the true cluster, and the y‐axis shows the fraction of regions predicted to belong to the clusters from Figure 3D (colors). ( D ) Average ChIP-seq signal of KLF4, KLF5, CHD4, SALL4, NANOG, ESRRB, MBD3, SOX2, DAX1, and TBX3 in ES cells 2 kb around regions in the clusters from Figure 3D . ( E ) Average ChIP-seq signal of CTCF and RAD21 in ES cells 2 kb around regions in the clusters from Figure 3D . Statistics for ( D–E ) are available in Supplementary file 1 .
    Figure Legend Snippet: Correlation between OCT4 binding and chromatin accessiblity, and analysis of results from random forest model. ( A ) Correlation between the log of normalized OCT4 ChIP-seq reads per bp (x-axis) and the log2 fold-change values of accessibility loss upon OCT4 depletion (y-axis) at all OCT4 binding sites in ZHBTc4 cells. ( B ) Correlation between the log of normalized OCT4 ChIP-seq reads per bp (x-axis) and the log of normalized ATAC-seq reads per bp (y-axis) at all OCT4 binding sites in ZHBTc4 cells. Coefficient (R) and p-values are based on the Pearson correlation coefficient. ( C ) Cluster predictions of regions in the test data based on a random forest model using mouse ES cell ChIP-seq data (see Materials and methods). The x‐axis shows the true cluster, and the y‐axis shows the fraction of regions predicted to belong to the clusters from Figure 3D (colors). ( D ) Average ChIP-seq signal of KLF4, KLF5, CHD4, SALL4, NANOG, ESRRB, MBD3, SOX2, DAX1, and TBX3 in ES cells 2 kb around regions in the clusters from Figure 3D . ( E ) Average ChIP-seq signal of CTCF and RAD21 in ES cells 2 kb around regions in the clusters from Figure 3D . Statistics for ( D–E ) are available in Supplementary file 1 .

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation

    Additional analyses of accessibility and binding changes upon SOX2 and OCT4 depletion. ( A ) Correlation between log2 fold-change values of accessibility (x-axis) and OCT4 binding (y-axis) upon SOX2 depletion in 2TS22C cells with dox treatment for 26 hr. Coefficient (R) and p-value are based on the Pearson correlation coefficient. ( B–C ) Average RPKM-normalized ATAC-seq signal 2 kb around OD (n = 3’730), CD (n = 1’463), and SD (n = 273) loci that overlap with a canonical OCT4::SOX2 motif upon SOX2 ( B ) and OCT4 ( C ) depletion. ( D–E ) Average RPKM-normalized OCT4 ( D ) and SOX2 ( E ) ChIP-seq signal 2 kb around OD, CD, and SD loci that overlap with a canonical OCT4::SOX2 motif upon SOX2 ( D ) and OCT4 ( E ) depletion. ( F ) Average RPKM-normalized ATAC-seq signal upon SOX2 depletion 2 kb around loci that display a significant increase in accessibility and SOX2 binding upon OCT4 depletion (n = 3’270). ( G ) Average RPKM-normalized BRG1 ChIP-seq signal upon OCT4 depletion 2 kb around loci that display a significant increase in accessibility and SOX2 binding upon OCT4 depletion. Statistics for ( B–G ) are available in Supplementary file 1 .
    Figure Legend Snippet: Additional analyses of accessibility and binding changes upon SOX2 and OCT4 depletion. ( A ) Correlation between log2 fold-change values of accessibility (x-axis) and OCT4 binding (y-axis) upon SOX2 depletion in 2TS22C cells with dox treatment for 26 hr. Coefficient (R) and p-value are based on the Pearson correlation coefficient. ( B–C ) Average RPKM-normalized ATAC-seq signal 2 kb around OD (n = 3’730), CD (n = 1’463), and SD (n = 273) loci that overlap with a canonical OCT4::SOX2 motif upon SOX2 ( B ) and OCT4 ( C ) depletion. ( D–E ) Average RPKM-normalized OCT4 ( D ) and SOX2 ( E ) ChIP-seq signal 2 kb around OD, CD, and SD loci that overlap with a canonical OCT4::SOX2 motif upon SOX2 ( D ) and OCT4 ( E ) depletion. ( F ) Average RPKM-normalized ATAC-seq signal upon SOX2 depletion 2 kb around loci that display a significant increase in accessibility and SOX2 binding upon OCT4 depletion (n = 3’270). ( G ) Average RPKM-normalized BRG1 ChIP-seq signal upon OCT4 depletion 2 kb around loci that display a significant increase in accessibility and SOX2 binding upon OCT4 depletion. Statistics for ( B–G ) are available in Supplementary file 1 .

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation

    Classification of OCT4/SOX2 binding sites. ( A ) Classification of all OCT4 and SOX2 binding sites into OD, CD, and SD loci as well as loci that were discarded due to differences in untreated cells between conditions or cell lines (Discarded), due to incongruent effect on accessibility after depletion in different conditions (Incongruent), and those that were increased in accessibility or unaffected by depletion. ( B ) ChromHMM signal enrichment at OD, CD, and SD loci as well as bound loci that were unaffected by depletion and loci not bound by OCT4 or SOX2.
    Figure Legend Snippet: Classification of OCT4/SOX2 binding sites. ( A ) Classification of all OCT4 and SOX2 binding sites into OD, CD, and SD loci as well as loci that were discarded due to differences in untreated cells between conditions or cell lines (Discarded), due to incongruent effect on accessibility after depletion in different conditions (Incongruent), and those that were increased in accessibility or unaffected by depletion. ( B ) ChromHMM signal enrichment at OD, CD, and SD loci as well as bound loci that were unaffected by depletion and loci not bound by OCT4 or SOX2.

    Techniques Used: Binding Assay

    Additional characterization of OD, CD, and SD loci. ( A ) Average ATAC-seq signal 2 kb around OD, CD, and SD loci in BRG1fl cells that were treated with tamoxifen (TAM) or left untreated. ( B ) Frequency of overlap (bar) and enrichment p-values (white digits) of the AP-2 motif 2 kb around OD, CD, and SD loci, and in background regions (BG). ( C ) Average ATAC-seq signal in TS cells 2 kb around OD, CD, and SD loci. ( D ) Average SOX2 ChIP-seq signal in TS cells 2 kb around OD, CD, and SD loci. ( E ) Percentage of the closest gene in the OD, CD, and SD groups as well as all other accessible regions (Other) whose nascent RNA levels are downregulated or upregulated upon 24 hr of OCT4 depletion. p-values: Fisher’s exact test. ( F ) Average ChIP-seq signal of ESRRB, NANOG, KLF4, and SALL4 in ES cells 2 kb around OD, CD, and SD loci. ( G ) Relative enrichment values (bar) and p-values (white digits) for the closest genes in the OD, CD, and SD groups, as well as loci upregulated upon SOX2 and OCT4 depletion, in the ‘Cell differentiation’ gene ontology set. ( H ) Average ChIP-seq signal of SOX2 2 kb around OD, CD, and SD loci in wt and PARP1 KO ES cells. Statistics for ( A ), ( C–D ), ( F ), ( H ) are available in Supplementary file 1 .
    Figure Legend Snippet: Additional characterization of OD, CD, and SD loci. ( A ) Average ATAC-seq signal 2 kb around OD, CD, and SD loci in BRG1fl cells that were treated with tamoxifen (TAM) or left untreated. ( B ) Frequency of overlap (bar) and enrichment p-values (white digits) of the AP-2 motif 2 kb around OD, CD, and SD loci, and in background regions (BG). ( C ) Average ATAC-seq signal in TS cells 2 kb around OD, CD, and SD loci. ( D ) Average SOX2 ChIP-seq signal in TS cells 2 kb around OD, CD, and SD loci. ( E ) Percentage of the closest gene in the OD, CD, and SD groups as well as all other accessible regions (Other) whose nascent RNA levels are downregulated or upregulated upon 24 hr of OCT4 depletion. p-values: Fisher’s exact test. ( F ) Average ChIP-seq signal of ESRRB, NANOG, KLF4, and SALL4 in ES cells 2 kb around OD, CD, and SD loci. ( G ) Relative enrichment values (bar) and p-values (white digits) for the closest genes in the OD, CD, and SD groups, as well as loci upregulated upon SOX2 and OCT4 depletion, in the ‘Cell differentiation’ gene ontology set. ( H ) Average ChIP-seq signal of SOX2 2 kb around OD, CD, and SD loci in wt and PARP1 KO ES cells. Statistics for ( A ), ( C–D ), ( F ), ( H ) are available in Supplementary file 1 .

    Techniques Used: Chromatin Immunoprecipitation, Cell Differentiation

    Immunofluorescence analysis of OCT4 OFF and SOX2 OFF cell lines and comparison of ATAC-seq changes between culture conditions and treatment times. ( A ) Immunofluorescence of 2TS22C cells stained for DNA (DAPI), OCT4, and SOX2 without dox treatment (left), and after 26 hr (middle), and 40 hr (right) of dox treatment. ( B ) Violin plot of background-subtracted log values of immunofluorescence signal in OCT4 (left) and SOX2 (right) channels upon SOX2 depletion. Control: n = 45’601 cells from four biological replicates including two technical replicates; 26 hr: n = 42’298 cells from three biological replicates including two technical replicates; 40 hr: n = 32’342 cells from two technical replicates. Dots: mean; Vertical lines: standard deviation; p-values: Mann-Whitney U. ( C ) Immunofluorescence of ZHBTc4 cells stained for DNA (DAPI), OCT4, and SOX2 without dox treatment (left), and after 24 hr of dox treatment (right). ( D ) Violin plot of background-subtracted log values of immunofluorescence signal in OCT4 (left) and SOX2 (right) channels upon OCT4 depletion. Control: n = 26’119 cells from three biological replicates. 24 hr: n = 23’157 cells from three biological replicates. Dots: mean; Vertical lines: standard deviation; p-values: Mann-Whitney U. ( E ) Correlation between the log2 fold-change values of accessibility upon OCT4 depletion in S2iL (x-axis) and SL (y-axis) at OCT4-bound sites. ( F ) Correlation between the log2 fold-change values of accessibility upon SOX2 depletion after 26 hr (x-axis) and 40 hr (y-axis) of dox treatment at SOX2 binding sites. Coefficient (R) and p-values are based on the Pearson correlation coefficient. Scale bars: 30 mm.
    Figure Legend Snippet: Immunofluorescence analysis of OCT4 OFF and SOX2 OFF cell lines and comparison of ATAC-seq changes between culture conditions and treatment times. ( A ) Immunofluorescence of 2TS22C cells stained for DNA (DAPI), OCT4, and SOX2 without dox treatment (left), and after 26 hr (middle), and 40 hr (right) of dox treatment. ( B ) Violin plot of background-subtracted log values of immunofluorescence signal in OCT4 (left) and SOX2 (right) channels upon SOX2 depletion. Control: n = 45’601 cells from four biological replicates including two technical replicates; 26 hr: n = 42’298 cells from three biological replicates including two technical replicates; 40 hr: n = 32’342 cells from two technical replicates. Dots: mean; Vertical lines: standard deviation; p-values: Mann-Whitney U. ( C ) Immunofluorescence of ZHBTc4 cells stained for DNA (DAPI), OCT4, and SOX2 without dox treatment (left), and after 24 hr of dox treatment (right). ( D ) Violin plot of background-subtracted log values of immunofluorescence signal in OCT4 (left) and SOX2 (right) channels upon OCT4 depletion. Control: n = 26’119 cells from three biological replicates. 24 hr: n = 23’157 cells from three biological replicates. Dots: mean; Vertical lines: standard deviation; p-values: Mann-Whitney U. ( E ) Correlation between the log2 fold-change values of accessibility upon OCT4 depletion in S2iL (x-axis) and SL (y-axis) at OCT4-bound sites. ( F ) Correlation between the log2 fold-change values of accessibility upon SOX2 depletion after 26 hr (x-axis) and 40 hr (y-axis) of dox treatment at SOX2 binding sites. Coefficient (R) and p-values are based on the Pearson correlation coefficient. Scale bars: 30 mm.

    Techniques Used: Immunofluorescence, Staining, Standard Deviation, MANN-WHITNEY, Binding Assay

    Heatmaps of ATAC-seq and ChIP-seq profiles in OCT4 OFF and SOX2 OFF cell lines at affected loci. Heatmaps of RPKM-normalized ATAC-seq and ChIP-seq binding profiles upon OCT4 ( A ) and SOX2 ( B ) depletion 5 kb around OCT4-regulated ( A ) and SOX2-regulated ( B ) loci. Each row represents one individual locus and each column represents one experimental condition.
    Figure Legend Snippet: Heatmaps of ATAC-seq and ChIP-seq profiles in OCT4 OFF and SOX2 OFF cell lines at affected loci. Heatmaps of RPKM-normalized ATAC-seq and ChIP-seq binding profiles upon OCT4 ( A ) and SOX2 ( B ) depletion 5 kb around OCT4-regulated ( A ) and SOX2-regulated ( B ) loci. Each row represents one individual locus and each column represents one experimental condition.

    Techniques Used: Chromatin Immunoprecipitation, Binding Assay

    5) Product Images from "Low P16INK4A Expression Associated with High Expression of Cancer Stem Cell Markers Predicts Poor Prognosis in Cervical Cancer after Radiotherapy"

    Article Title: Low P16INK4A Expression Associated with High Expression of Cancer Stem Cell Markers Predicts Poor Prognosis in Cervical Cancer after Radiotherapy

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19092541

    The inhibition of P16 INK4A protein expression increased SOX2, ALDH1A1 expression, and self-renewal ability in cervical cancer cells. ( A ) Expression of stem cell markers (CD133, SOX2, and ALDH1A1) at the protein level in the HeLa-control and HeLa-shP16 cells were detected by Western blot and relative protein expression to beta-actin protein was shown. ( B ) Colony formation assay of HeLa-control and HeLa-shP16 cells were used to evaluate the ability of self-renew. ( C ) The sphere formation assay was used to measure the anchorage-independent growth of HeLa-control and HeLa-shP16 cells in ultra-low attachment plates. Each bar represents the mean ± SD of three independent experiments. Student’s t -test was used for continuous variables between the two groups. * p
    Figure Legend Snippet: The inhibition of P16 INK4A protein expression increased SOX2, ALDH1A1 expression, and self-renewal ability in cervical cancer cells. ( A ) Expression of stem cell markers (CD133, SOX2, and ALDH1A1) at the protein level in the HeLa-control and HeLa-shP16 cells were detected by Western blot and relative protein expression to beta-actin protein was shown. ( B ) Colony formation assay of HeLa-control and HeLa-shP16 cells were used to evaluate the ability of self-renew. ( C ) The sphere formation assay was used to measure the anchorage-independent growth of HeLa-control and HeLa-shP16 cells in ultra-low attachment plates. Each bar represents the mean ± SD of three independent experiments. Student’s t -test was used for continuous variables between the two groups. * p

    Techniques Used: Inhibition, Expressing, Western Blot, Colony Assay, Tube Formation Assay

    Survival and recurrence outcomes of patients with different expressions of P16 INK4A , SOX2, and ALDH1A1 in tumors. ( A , B ) Cervical cancer patients with high P16 INK4A expression had a better five-year OS rate ( p = 0.016) and better five-year DFS rate ( p = 0.02) than those with lower expression. ( C , D ) Patients with high SOX2 expression had similar five-year OS and DFS than those with low expression ( C , p = 0.598 and D , p = 0.141). ( E , F ) Patients with high ALDH1A1 expression had similar five-year OS and DFS than those with low expression ( E , p = 0.591 and F , p = 0.131). ( G , H ) The patients with low P16 INK4A /high SOX2 expression had similar five-year OS rates ( G , p = 0.118) but worse five-year DFS rates ( H , p = 0.009) than those with high P16 INK4A /lower SOX2 expression. ( I , J ) The patients with low P16 INK4A /high ALDH1A1 expression had worse five-year OS rates ( I , p = 0.030) and worse five-year DFS rates ( J , p = 0.003) than those with high P16 INK4A /lower ALDH1A1 expression.
    Figure Legend Snippet: Survival and recurrence outcomes of patients with different expressions of P16 INK4A , SOX2, and ALDH1A1 in tumors. ( A , B ) Cervical cancer patients with high P16 INK4A expression had a better five-year OS rate ( p = 0.016) and better five-year DFS rate ( p = 0.02) than those with lower expression. ( C , D ) Patients with high SOX2 expression had similar five-year OS and DFS than those with low expression ( C , p = 0.598 and D , p = 0.141). ( E , F ) Patients with high ALDH1A1 expression had similar five-year OS and DFS than those with low expression ( E , p = 0.591 and F , p = 0.131). ( G , H ) The patients with low P16 INK4A /high SOX2 expression had similar five-year OS rates ( G , p = 0.118) but worse five-year DFS rates ( H , p = 0.009) than those with high P16 INK4A /lower SOX2 expression. ( I , J ) The patients with low P16 INK4A /high ALDH1A1 expression had worse five-year OS rates ( I , p = 0.030) and worse five-year DFS rates ( J , p = 0.003) than those with high P16 INK4A /lower ALDH1A1 expression.

    Techniques Used: Expressing

    Immunostaining of P16 INK4A , SOX2, and ALDH1A1 expression in pretreatment cervical cancer. Immunohistochemical staining of P16 INK4A expression was low in ( A ) and high in ( D ), SOX2 expression was low in ( B ) and high in ( E ), and ALDH1A1 expression was low in ( C ) and high in ( F ). Scale bar: 100 μm.
    Figure Legend Snippet: Immunostaining of P16 INK4A , SOX2, and ALDH1A1 expression in pretreatment cervical cancer. Immunohistochemical staining of P16 INK4A expression was low in ( A ) and high in ( D ), SOX2 expression was low in ( B ) and high in ( E ), and ALDH1A1 expression was low in ( C ) and high in ( F ). Scale bar: 100 μm.

    Techniques Used: Immunostaining, Expressing, Immunohistochemistry, Staining

    6) Product Images from "Low P16INK4A Expression Associated with High Expression of Cancer Stem Cell Markers Predicts Poor Prognosis in Cervical Cancer after Radiotherapy"

    Article Title: Low P16INK4A Expression Associated with High Expression of Cancer Stem Cell Markers Predicts Poor Prognosis in Cervical Cancer after Radiotherapy

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19092541

    The inhibition of P16 INK4A protein expression increased SOX2, ALDH1A1 expression, and self-renewal ability in cervical cancer cells. ( A ) Expression of stem cell markers (CD133, SOX2, and ALDH1A1) at the protein level in the HeLa-control and HeLa-shP16 cells were detected by Western blot and relative protein expression to beta-actin protein was shown. ( B ) Colony formation assay of HeLa-control and HeLa-shP16 cells were used to evaluate the ability of self-renew. ( C ) The sphere formation assay was used to measure the anchorage-independent growth of HeLa-control and HeLa-shP16 cells in ultra-low attachment plates. Each bar represents the mean ± SD of three independent experiments. Student’s t -test was used for continuous variables between the two groups. * p
    Figure Legend Snippet: The inhibition of P16 INK4A protein expression increased SOX2, ALDH1A1 expression, and self-renewal ability in cervical cancer cells. ( A ) Expression of stem cell markers (CD133, SOX2, and ALDH1A1) at the protein level in the HeLa-control and HeLa-shP16 cells were detected by Western blot and relative protein expression to beta-actin protein was shown. ( B ) Colony formation assay of HeLa-control and HeLa-shP16 cells were used to evaluate the ability of self-renew. ( C ) The sphere formation assay was used to measure the anchorage-independent growth of HeLa-control and HeLa-shP16 cells in ultra-low attachment plates. Each bar represents the mean ± SD of three independent experiments. Student’s t -test was used for continuous variables between the two groups. * p

    Techniques Used: Inhibition, Expressing, Western Blot, Colony Assay, Tube Formation Assay

    7) Product Images from "Cancer Stem Cells in Glioblastoma Multiforme"

    Article Title: Cancer Stem Cells in Glioblastoma Multiforme

    Journal: Frontiers in Surgery

    doi: 10.3389/fsurg.2016.00048

    Representative images of IF IHC-stained sections of GBM tissue for ESC markers . SOX2 [ (A) , red, arrows ], NANOG [ (B) , red, arrows ], and pSTAT3 [ (C) , red, arrows ] all showed nuclear expression on GFAP+ tumor cells [ (A–C) , green]. OCT4 [ (D) , red, arrows ] staining was scarce and solely cytoplasmic in GFAP+ tumor cells. SALL4 [ (E) , green] and SOX2 [ (E) , red] were co-expressed ( arrows ) in the nuclei of some tumor cells, with SALL4 also staining SOX2–negative cells. Cell nuclei were counterstained with 4′, 6′-diamidino-2-phenylindole [(A–E), blue]. Scale bars: 20 μm.
    Figure Legend Snippet: Representative images of IF IHC-stained sections of GBM tissue for ESC markers . SOX2 [ (A) , red, arrows ], NANOG [ (B) , red, arrows ], and pSTAT3 [ (C) , red, arrows ] all showed nuclear expression on GFAP+ tumor cells [ (A–C) , green]. OCT4 [ (D) , red, arrows ] staining was scarce and solely cytoplasmic in GFAP+ tumor cells. SALL4 [ (E) , green] and SOX2 [ (E) , red] were co-expressed ( arrows ) in the nuclei of some tumor cells, with SALL4 also staining SOX2–negative cells. Cell nuclei were counterstained with 4′, 6′-diamidino-2-phenylindole [(A–E), blue]. Scale bars: 20 μm.

    Techniques Used: Immunohistochemistry, Staining, Expressing

    Representative image of H E stained slides showing the presence of GBM (A) and DAB IHC-stained slides of GBM tissue demonstrating expression of CSC markers (B–F) . pSTAT3 [ (B) , brown] was expressed in the nuclei of tumor and endothelium of the microvessels throughout the sample. SALL4 [ (C) , brown] was predominantly expressed on the nuclei of the tumor cells, and the cytoplasm of the endothelial cells lining the microvessels. SOX2 [ (D) , brown] displayed strong nuclear staining of the tumor cells, and moderate cytoplasmic staining of the endothelium of the microvessels. Nuclear expression of NANOG [ (E) , pink/red] was observed on the tumor cells, and to the lesser extent on the endothelium of the microvessels. Nuclear and cytoplasmic staining of OCT4 [ (F) , brown] was observed in few tumor cells. Cell nuclei were counterstained with hematoxylin [ (A–F) , blue]. Original magnification: 400X.
    Figure Legend Snippet: Representative image of H E stained slides showing the presence of GBM (A) and DAB IHC-stained slides of GBM tissue demonstrating expression of CSC markers (B–F) . pSTAT3 [ (B) , brown] was expressed in the nuclei of tumor and endothelium of the microvessels throughout the sample. SALL4 [ (C) , brown] was predominantly expressed on the nuclei of the tumor cells, and the cytoplasm of the endothelial cells lining the microvessels. SOX2 [ (D) , brown] displayed strong nuclear staining of the tumor cells, and moderate cytoplasmic staining of the endothelium of the microvessels. Nuclear expression of NANOG [ (E) , pink/red] was observed on the tumor cells, and to the lesser extent on the endothelium of the microvessels. Nuclear and cytoplasmic staining of OCT4 [ (F) , brown] was observed in few tumor cells. Cell nuclei were counterstained with hematoxylin [ (A–F) , blue]. Original magnification: 400X.

    Techniques Used: Staining, Immunohistochemistry, Expressing

    Western blots of five GBM tissue samples . OCT4 was not detected (A) , pSTAT3 at ~90 kDa was found in three out of five samples (B) , NANOG was present in all five samples with multiple bands at approximately 40 and 31 kDa (C) . SOX2 was detected in four out of five samples with bands at approximately 45 and 38 kDa (D) .
    Figure Legend Snippet: Western blots of five GBM tissue samples . OCT4 was not detected (A) , pSTAT3 at ~90 kDa was found in three out of five samples (B) , NANOG was present in all five samples with multiple bands at approximately 40 and 31 kDa (C) . SOX2 was detected in four out of five samples with bands at approximately 45 and 38 kDa (D) .

    Techniques Used: Western Blot

    Relative expression of CSC mRNA transcripts in six GBM samples showing the presence of all 5 markers at varying levels . NANOG, OCT4, and SALL4 showed relatively low mRNA expression, while STAT3 and SOX2 displayed high levels of mRNA expression. Expression is depicted relative to the housekeeper GUSB.
    Figure Legend Snippet: Relative expression of CSC mRNA transcripts in six GBM samples showing the presence of all 5 markers at varying levels . NANOG, OCT4, and SALL4 showed relatively low mRNA expression, while STAT3 and SOX2 displayed high levels of mRNA expression. Expression is depicted relative to the housekeeper GUSB.

    Techniques Used: Expressing

    8) Product Images from "Autophagy and Cellular Senescence Mediated by Sox2 Suppress Malignancy of Cancer Cells"

    Article Title: Autophagy and Cellular Senescence Mediated by Sox2 Suppress Malignancy of Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0057172

    Sox2-induced autophagy suppresses cancer growth in a xenograft mouse model. ( A ) Xenograft in vivo tumor growth. HCT116 cells (3×10 6 ) stably expressing mock (n = 9) or Sox2 (n = 10) were injected subcutaneously into the dorsal right flank of athymic nude mice. At the endpoint (30 days) after injection, mice were euthanized and necropsied. ( B ) Sox2 expression delayed tumor growth in athymic nude mice. Tumor incidence of HCT116 cells stably expressing mock or Sox2 was analyzed at first measurable tumor growth after injection and compared (*p
    Figure Legend Snippet: Sox2-induced autophagy suppresses cancer growth in a xenograft mouse model. ( A ) Xenograft in vivo tumor growth. HCT116 cells (3×10 6 ) stably expressing mock (n = 9) or Sox2 (n = 10) were injected subcutaneously into the dorsal right flank of athymic nude mice. At the endpoint (30 days) after injection, mice were euthanized and necropsied. ( B ) Sox2 expression delayed tumor growth in athymic nude mice. Tumor incidence of HCT116 cells stably expressing mock or Sox2 was analyzed at first measurable tumor growth after injection and compared (*p

    Techniques Used: In Vivo, Stable Transfection, Expressing, Injection, Mouse Assay

    Sox2-induced autophagy is mediated through the down-regulation of the Akt signaling pathway, but not through Class III PI3-K signaling. ( A–C ) Protein levels of Class I signaling molecules including Akt, mTOR and p70S6K and class III PI3-K signaling molecules including Vps34p and beclin were analyzed in HCT116 cells stably expressing mock or Sox2 . The individual proteins were visualized by Western blotting using specific antibodies. β-Actin was used as an internal control to verify equal protein loading. ( D ) Involvement of Class I PI3-K signaling in Sox2-induced autophagy. ( D, left panels ) HCT116 cells stably expressing mock or Sox2 were treated with insulin or LY294002 under 10% FBS-containing conditions at 2 days after infection. Vacuole formation was observed by light microscopy (X200). The boxed areas are individually magnified 2 more times ( lower panels for mock and Sox2 ) to better visualize vacuoles. (D, right panels) The cells were transduced with Sox2 viral particles and cultured 2 days. The cell culture medium was replaced with McCoy’s 5a without FBS supplementation but including 5 µg/ml insulin or 5 µg LY294002 and then cells were cultured for 5 days. The cells were observed under light microscopy (X200). ( E ) Quantitative comparison of autophagy formation induced by class I PI3-K signaling. The vacuole forming cells from D and F were counted and compared in a numerate graphic (*p
    Figure Legend Snippet: Sox2-induced autophagy is mediated through the down-regulation of the Akt signaling pathway, but not through Class III PI3-K signaling. ( A–C ) Protein levels of Class I signaling molecules including Akt, mTOR and p70S6K and class III PI3-K signaling molecules including Vps34p and beclin were analyzed in HCT116 cells stably expressing mock or Sox2 . The individual proteins were visualized by Western blotting using specific antibodies. β-Actin was used as an internal control to verify equal protein loading. ( D ) Involvement of Class I PI3-K signaling in Sox2-induced autophagy. ( D, left panels ) HCT116 cells stably expressing mock or Sox2 were treated with insulin or LY294002 under 10% FBS-containing conditions at 2 days after infection. Vacuole formation was observed by light microscopy (X200). The boxed areas are individually magnified 2 more times ( lower panels for mock and Sox2 ) to better visualize vacuoles. (D, right panels) The cells were transduced with Sox2 viral particles and cultured 2 days. The cell culture medium was replaced with McCoy’s 5a without FBS supplementation but including 5 µg/ml insulin or 5 µg LY294002 and then cells were cultured for 5 days. The cells were observed under light microscopy (X200). ( E ) Quantitative comparison of autophagy formation induced by class I PI3-K signaling. The vacuole forming cells from D and F were counted and compared in a numerate graphic (*p

    Techniques Used: Stable Transfection, Expressing, Western Blot, Infection, Light Microscopy, Transduction, Cell Culture

    Sox2-induced autophagy causes senescence in HCT116 colorectal cancer cells resulting in suppressed proliferation and decreased anchorage-independent colony growth. ( A ) Left panel , effect of Sox2 expression on cell proliferation. HCT116 cells (2×10 3 ) stably expressing mock or Sox2 were seeded in 96-well plates and proliferation was analyzed by MTS assay at 24 h intervals up to 96 h (*, p
    Figure Legend Snippet: Sox2-induced autophagy causes senescence in HCT116 colorectal cancer cells resulting in suppressed proliferation and decreased anchorage-independent colony growth. ( A ) Left panel , effect of Sox2 expression on cell proliferation. HCT116 cells (2×10 3 ) stably expressing mock or Sox2 were seeded in 96-well plates and proliferation was analyzed by MTS assay at 24 h intervals up to 96 h (*, p

    Techniques Used: Expressing, Stable Transfection, MTS Assay

    Sox2 targets ATG10 to induce autophagy. ( A ) Microarray. Top panel , HCT116 colorectal cancer cells stably expressing mock or Sox2 were subjected to microarray analysis as described in “Materials and Methods”. The yellow color indicates the number of genes that did not show a significant difference between mock and Sox2 expression. The red and green colors indicate the number of genes up- or down-regulated, respectively, in HCT116 cells stably expressing Sox2 compared with cells expressing the mock control. Bottom table , summary of autophagy-related genes obtained from the microarray analysis. Up- and down-regulation are denoted in Log2 values. ( B ) Expression of ATG10 induced by Sox2. Total RNA (1 µg) from HCT116 cells infected with mock or Sox2 was reverse transcribed and ATG10 was amplified by cycle-dependent PCR and the ATG10 expression level was visualized by agarose gel electrophoresis at the 21 st cycle. ß-Actin was used as an internal control to verify equal utilization of cDNA for PCR. ( C ) Up-regulation of ATG10 and LC3b protein levels induced by Sox2 expression. Proteins were extracted from HCT116 cells stably expressing mock or Sox2 and ATG10 and LC3b proteins were visualized by Western blotting using specific antibodies. ß-Actin was used as an internal control to verify equal protein loading. ( D ) Confirmation of Sox2-induced ATG10 protein level. HCT116 colorectal cancer cells were infected with mock or Sox2 and cultured 5 days. The cells were fixed, hybridized with an ATG10 specific primary antibody and Alexa 488-conjugated secondary antibody. The ATG10 protein levels were observed by fluorescence microscopy. L.M. indicates the same area of light microscopy corresponding to fluorescence microscopy (X200). ( E ) Construction of the ATG10 promoter luciferase reporter plasmid. The nucleotide sequences of the human ATG10 promoter region were downloaded from Ensemble ( http://uswest.ensemble.org ). The putative promoter analysis was conducted using TFSEARCH (v1.3) ( http://cbrc.jp/htbin/nph-tfsearch ). The putative SRY and Sox binding consensus sequences are denoted in red and the polymerase III binding site is boxed. ( F ) The BAC clone (RP11-111B20) was purchased from Empire Genomics (Buffalo, NY) and 1528 and 711 bp of the ATG10 promoter region were amplified by PCR. The PCR fragment was recombined with the pGL3 basic vector to construct pGL3-AT10-1582 and pGL3-ATG10-711 luciferase reporter plasmids. The pGL3-ATG10-luc reporter plasmids were confirmed by DNA sequencing. The ATG10 promoter luciferase reporter plasmids, pGL3-AT10-1582 and pGL3-ATG10-71- luc, were transiently transfected into HCT116 cells and the firefly luciferase activity was analyzed after 24 h. The phRL-SV40 renilla luciferase reporter plasmid was co-transfected as an internal control to verify equal transfection and normalization of firefly luciferase activity (*p
    Figure Legend Snippet: Sox2 targets ATG10 to induce autophagy. ( A ) Microarray. Top panel , HCT116 colorectal cancer cells stably expressing mock or Sox2 were subjected to microarray analysis as described in “Materials and Methods”. The yellow color indicates the number of genes that did not show a significant difference between mock and Sox2 expression. The red and green colors indicate the number of genes up- or down-regulated, respectively, in HCT116 cells stably expressing Sox2 compared with cells expressing the mock control. Bottom table , summary of autophagy-related genes obtained from the microarray analysis. Up- and down-regulation are denoted in Log2 values. ( B ) Expression of ATG10 induced by Sox2. Total RNA (1 µg) from HCT116 cells infected with mock or Sox2 was reverse transcribed and ATG10 was amplified by cycle-dependent PCR and the ATG10 expression level was visualized by agarose gel electrophoresis at the 21 st cycle. ß-Actin was used as an internal control to verify equal utilization of cDNA for PCR. ( C ) Up-regulation of ATG10 and LC3b protein levels induced by Sox2 expression. Proteins were extracted from HCT116 cells stably expressing mock or Sox2 and ATG10 and LC3b proteins were visualized by Western blotting using specific antibodies. ß-Actin was used as an internal control to verify equal protein loading. ( D ) Confirmation of Sox2-induced ATG10 protein level. HCT116 colorectal cancer cells were infected with mock or Sox2 and cultured 5 days. The cells were fixed, hybridized with an ATG10 specific primary antibody and Alexa 488-conjugated secondary antibody. The ATG10 protein levels were observed by fluorescence microscopy. L.M. indicates the same area of light microscopy corresponding to fluorescence microscopy (X200). ( E ) Construction of the ATG10 promoter luciferase reporter plasmid. The nucleotide sequences of the human ATG10 promoter region were downloaded from Ensemble ( http://uswest.ensemble.org ). The putative promoter analysis was conducted using TFSEARCH (v1.3) ( http://cbrc.jp/htbin/nph-tfsearch ). The putative SRY and Sox binding consensus sequences are denoted in red and the polymerase III binding site is boxed. ( F ) The BAC clone (RP11-111B20) was purchased from Empire Genomics (Buffalo, NY) and 1528 and 711 bp of the ATG10 promoter region were amplified by PCR. The PCR fragment was recombined with the pGL3 basic vector to construct pGL3-AT10-1582 and pGL3-ATG10-711 luciferase reporter plasmids. The pGL3-ATG10-luc reporter plasmids were confirmed by DNA sequencing. The ATG10 promoter luciferase reporter plasmids, pGL3-AT10-1582 and pGL3-ATG10-71- luc, were transiently transfected into HCT116 cells and the firefly luciferase activity was analyzed after 24 h. The phRL-SV40 renilla luciferase reporter plasmid was co-transfected as an internal control to verify equal transfection and normalization of firefly luciferase activity (*p

    Techniques Used: Microarray, Stable Transfection, Expressing, Infection, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Western Blot, Cell Culture, Fluorescence, Microscopy, Light Microscopy, Luciferase, Plasmid Preparation, Binding Assay, BAC Assay, Empire Assay, Construct, DNA Sequencing, Transfection, Activity Assay

    Ectopic expression of Sox2 induces autophagy. ( A ) Comparison of morphological changes induced by ectopic expression of iPS factors. (Upper panels) HCT116 cells were individually transduced with iPS factors, including Sox2, Nanog, Lin28 and Oct4. The cells were cultured for 5 days and changes were observed under a light microscope (X200). (Lower panels) HCT116 cells were harvested at 5 days after transduction with iPS factors and proteins extracted. The protein levels of the Sox2, Nanog, Lin28 and Oct4 were analyzed by Western blotting with specific antibodies as indicated. ß-Actin was used as an internal control to verify equal protein loading. ( B ) HCT116 cells forming vacuoles at 5 days after transduction were observed under light microscopy, counted and compared. Sox2 overexpression was analyzed by Western blotting and immunocytofluorescence assay (X200) using specific antibodies as indicated. ß-Actin was used as an internal control to verify equal protein loading. The cells were visualized by light microscopy (L.M.; X200). ( C ) Lysosomal activation analysis. HCT116 cells infected with mock or Sox2 were stained by adding lysotracker (50 nM) into the culture medium for 5 min in a 37 o C, 5% CO 2 incubator. The cells were fixed with 4% formalin, washed with PBS and lysosomal activation was observed under a fluorescence microscope (X200). ( D ) Immunofluorescence assay of LC3b (i.e., ATG8b). HCT116 cells stably expressing mock or Sox2 were subjected to a fluorescence assay to detect LC3b. The cells were observed under a fluorescence microscope (X200). The nuclei were stained with DAPI; L.M. indicates light microscopy (X200).
    Figure Legend Snippet: Ectopic expression of Sox2 induces autophagy. ( A ) Comparison of morphological changes induced by ectopic expression of iPS factors. (Upper panels) HCT116 cells were individually transduced with iPS factors, including Sox2, Nanog, Lin28 and Oct4. The cells were cultured for 5 days and changes were observed under a light microscope (X200). (Lower panels) HCT116 cells were harvested at 5 days after transduction with iPS factors and proteins extracted. The protein levels of the Sox2, Nanog, Lin28 and Oct4 were analyzed by Western blotting with specific antibodies as indicated. ß-Actin was used as an internal control to verify equal protein loading. ( B ) HCT116 cells forming vacuoles at 5 days after transduction were observed under light microscopy, counted and compared. Sox2 overexpression was analyzed by Western blotting and immunocytofluorescence assay (X200) using specific antibodies as indicated. ß-Actin was used as an internal control to verify equal protein loading. The cells were visualized by light microscopy (L.M.; X200). ( C ) Lysosomal activation analysis. HCT116 cells infected with mock or Sox2 were stained by adding lysotracker (50 nM) into the culture medium for 5 min in a 37 o C, 5% CO 2 incubator. The cells were fixed with 4% formalin, washed with PBS and lysosomal activation was observed under a fluorescence microscope (X200). ( D ) Immunofluorescence assay of LC3b (i.e., ATG8b). HCT116 cells stably expressing mock or Sox2 were subjected to a fluorescence assay to detect LC3b. The cells were observed under a fluorescence microscope (X200). The nuclei were stained with DAPI; L.M. indicates light microscopy (X200).

    Techniques Used: Expressing, Transduction, Cell Culture, Light Microscopy, Western Blot, Over Expression, Activation Assay, Infection, Staining, Fluorescence, Microscopy, Immunofluorescence, Stable Transfection

    Knockdown of ATG10 restores Sox2-induced autophagy, cellular senescence and proliferation. ( A ) Confirmation of the knockdown efficiency of ATG10. Knockdown of ATG10 using pLKO-shATG10 . The pLKO-shATG10 knockdown lenti-viral particles were infected into HCT116 cells stably expressing Sox2 and cells cultured for 36 h. The proteins were extracted and knockdown efficiency was determined by Western blot using a specific ATG10 antibody. ß-Actin was used as an internal control to verify equal protein loading. ( B ) Morphological changes induced by ATG10 knockdown in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells were analyzed for morphological changes and protein levels of Sox2 (red) and ATG10 (green) were determined by an immunofluorescence assay using specific antibodies. The cells were observed under a fluorescence or light microscope (X200). L.M. indicates light microscopy. ( C ) Restoration of cell cycle, cell proliferation and senescence markers by knocking down ATG10 in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells were seeded into a 4-chamber slide, fixed and permeabilized. Markers for cell proliferation, cellular senescence and cell cycle regulation included Ki-67, ß-galactosidase, p16 INK4a and p21. These markers were analyzed by immunocytochemistry with specific antibodies as indicated using the Sigma FASTTM 3,3′-Diaminobenzidine Terahyfrochloride with Metal Enhancer Tablet Sets (DAB peroxidase substrate). The cells were observed under a fluorescence or light microscope (X200). ( D ) Restoration of proliferation by knocking down ATG10 in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells (2×10 3 ) were seeded into 96-well plates and proliferation was analyzed by MTS assay at 24 h intervals up to 96 h (*p
    Figure Legend Snippet: Knockdown of ATG10 restores Sox2-induced autophagy, cellular senescence and proliferation. ( A ) Confirmation of the knockdown efficiency of ATG10. Knockdown of ATG10 using pLKO-shATG10 . The pLKO-shATG10 knockdown lenti-viral particles were infected into HCT116 cells stably expressing Sox2 and cells cultured for 36 h. The proteins were extracted and knockdown efficiency was determined by Western blot using a specific ATG10 antibody. ß-Actin was used as an internal control to verify equal protein loading. ( B ) Morphological changes induced by ATG10 knockdown in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells were analyzed for morphological changes and protein levels of Sox2 (red) and ATG10 (green) were determined by an immunofluorescence assay using specific antibodies. The cells were observed under a fluorescence or light microscope (X200). L.M. indicates light microscopy. ( C ) Restoration of cell cycle, cell proliferation and senescence markers by knocking down ATG10 in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells were seeded into a 4-chamber slide, fixed and permeabilized. Markers for cell proliferation, cellular senescence and cell cycle regulation included Ki-67, ß-galactosidase, p16 INK4a and p21. These markers were analyzed by immunocytochemistry with specific antibodies as indicated using the Sigma FASTTM 3,3′-Diaminobenzidine Terahyfrochloride with Metal Enhancer Tablet Sets (DAB peroxidase substrate). The cells were observed under a fluorescence or light microscope (X200). ( D ) Restoration of proliferation by knocking down ATG10 in HCT116 cells stably expressing Sox2 . HCT116-mock, -Sox2 and -Sox2/sh-ATG10 cells (2×10 3 ) were seeded into 96-well plates and proliferation was analyzed by MTS assay at 24 h intervals up to 96 h (*p

    Techniques Used: Infection, Stable Transfection, Expressing, Cell Culture, Western Blot, Immunofluorescence, Fluorescence, Light Microscopy, Immunocytochemistry, MTS Assay

    Cancer cell-specific lysosomal activation. ( A ) CCD-18Co (CRL-1459) normal colon cells and HCT116, HT29 and WiDr colon cancer cells were cultured in the appropriate medium and transduced with Sox2 viral particles. The cells were cultured with complete growth medium for 5 days after transduction. The cells were observed under a light microscope. Cellular vacuoles are marked with an arrowhead in Sox2 -expressing colon cancer cells. ( B ) CCD-18Co normal colon cells were cultured, transduced with mock or Sox2 viral particles and cultured with complete growth medium for 5 days. The cells were then fixed, permeabilized and hybridized with a Sox2 specific antibody and then with an Alexa 568-conjugated secondary antibody, and visualized with a fluorescence microscope (X200). Nuclei were stained with DAPI. The areas of light microscopy are the areas matched with Sox2 and DAPI fluorescence. The boxed area under low power is magnified to compare the morphology between HCT116- mock and - Sox2 -expressing cells. ( C ) Lysosomal activation, a autophagy marker, was compared by adding lysotracker-Red (50 nM) at 5 days after transduction to CCD-18Co normal colon cells and HCT116 colorectal cancer cells infected with mock or Sox2 . The cells were observed under a fluorescence microscope. L.M. indicates the same area of light microscopy corresponding to fluorescence microscopy (X200).
    Figure Legend Snippet: Cancer cell-specific lysosomal activation. ( A ) CCD-18Co (CRL-1459) normal colon cells and HCT116, HT29 and WiDr colon cancer cells were cultured in the appropriate medium and transduced with Sox2 viral particles. The cells were cultured with complete growth medium for 5 days after transduction. The cells were observed under a light microscope. Cellular vacuoles are marked with an arrowhead in Sox2 -expressing colon cancer cells. ( B ) CCD-18Co normal colon cells were cultured, transduced with mock or Sox2 viral particles and cultured with complete growth medium for 5 days. The cells were then fixed, permeabilized and hybridized with a Sox2 specific antibody and then with an Alexa 568-conjugated secondary antibody, and visualized with a fluorescence microscope (X200). Nuclei were stained with DAPI. The areas of light microscopy are the areas matched with Sox2 and DAPI fluorescence. The boxed area under low power is magnified to compare the morphology between HCT116- mock and - Sox2 -expressing cells. ( C ) Lysosomal activation, a autophagy marker, was compared by adding lysotracker-Red (50 nM) at 5 days after transduction to CCD-18Co normal colon cells and HCT116 colorectal cancer cells infected with mock or Sox2 . The cells were observed under a fluorescence microscope. L.M. indicates the same area of light microscopy corresponding to fluorescence microscopy (X200).

    Techniques Used: Activation Assay, Cell Culture, Transduction, Light Microscopy, Expressing, Fluorescence, Microscopy, Staining, Marker, Infection

    9) Product Images from "Sorting Live Stem Cells Based on Sox2 mRNA Expression"

    Article Title: Sorting Live Stem Cells Based on Sox2 mRNA Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0049874

    Isolation of neurospheres from primary mouse tissue and of in vitro cultured neurospheres using Sox2- MB. (A) Two cell populations, namely Sox2- MB high and the Sox2- MB low , were first selected on Annexin-V - cells and then by comparing the nonspecific-MB fluorescent signal to the Sox2-MB fluorescent signal. (B) After 1 wk, sphere-forming efficiency was calculated from the Sox2- MB high and the Sox2- MB low populations as well as non-sorted primary mouse hippocampus isolated cells. (C and D) Images of 1 wk old spheres generated from sorted Sox2- MB low cells and Sox2- MB high cells (scale bar = 25 µm). (E) Neurospheres from the Sox2- MB high and the Sox2- MB low populations were serially passaged and cumulative population doublings was calculated. (F) In vitro cultured neurosphere mRNA expression of Sox2 was analyzed by RT-PCR and compared to MEFs. (G) Two cell populations, namely Sox2 -MB high and Sox2- MB low , were selected by comparing the nonspecific-MB fluorescent signal to the Sox2- MB fluorescent signal. (H) After 1 wk, sphere-forming efficiencies were calculated. (I) Neurospheres formed by the Sox2- MB high and the Sox2- MB low populations were stained for Sox2 and Nestin, or secondary antibodies only (control) (scale bar = 50 µm). Error bars represent the mean ± SEM. Asterisks denotes statistical significance ((n = 5 samples * p
    Figure Legend Snippet: Isolation of neurospheres from primary mouse tissue and of in vitro cultured neurospheres using Sox2- MB. (A) Two cell populations, namely Sox2- MB high and the Sox2- MB low , were first selected on Annexin-V - cells and then by comparing the nonspecific-MB fluorescent signal to the Sox2-MB fluorescent signal. (B) After 1 wk, sphere-forming efficiency was calculated from the Sox2- MB high and the Sox2- MB low populations as well as non-sorted primary mouse hippocampus isolated cells. (C and D) Images of 1 wk old spheres generated from sorted Sox2- MB low cells and Sox2- MB high cells (scale bar = 25 µm). (E) Neurospheres from the Sox2- MB high and the Sox2- MB low populations were serially passaged and cumulative population doublings was calculated. (F) In vitro cultured neurosphere mRNA expression of Sox2 was analyzed by RT-PCR and compared to MEFs. (G) Two cell populations, namely Sox2 -MB high and Sox2- MB low , were selected by comparing the nonspecific-MB fluorescent signal to the Sox2- MB fluorescent signal. (H) After 1 wk, sphere-forming efficiencies were calculated. (I) Neurospheres formed by the Sox2- MB high and the Sox2- MB low populations were stained for Sox2 and Nestin, or secondary antibodies only (control) (scale bar = 50 µm). Error bars represent the mean ± SEM. Asterisks denotes statistical significance ((n = 5 samples * p

    Techniques Used: Isolation, In Vitro, Cell Culture, Generated, Expressing, Reverse Transcription Polymerase Chain Reaction, Staining

    Mechanism and design of Sox2 mRNA-specific MB. (A) Opening of the Sox2 -MB is induced in the cytoplasm of cells expressing Sox2 and emission of Cy3 fluorescence is detected. In contrast, Sox2 -MB remains in the hairpin conformation in the cytoplasm of Sox2 negative cells, and no emission of Cy3 fluorescence is detected. (B) The sequences of the designed Sox2 -MBs and the non-specific-MB. (C) The sequences of the synthesized oligonucleotides complementary to the loop sequence of each Sox2 -MB. (D) The Sox2 -MBs were mixed with or without its target sequence and the Cy3-fluorescence was detected with microplate reader. A difference was seen in all of the designed Sox2 -MBs between when the target sequence was present or not. Error bars represent the mean ± SEM. Asterisks denote statistical significance (n = 3 samples, *** p
    Figure Legend Snippet: Mechanism and design of Sox2 mRNA-specific MB. (A) Opening of the Sox2 -MB is induced in the cytoplasm of cells expressing Sox2 and emission of Cy3 fluorescence is detected. In contrast, Sox2 -MB remains in the hairpin conformation in the cytoplasm of Sox2 negative cells, and no emission of Cy3 fluorescence is detected. (B) The sequences of the designed Sox2 -MBs and the non-specific-MB. (C) The sequences of the synthesized oligonucleotides complementary to the loop sequence of each Sox2 -MB. (D) The Sox2 -MBs were mixed with or without its target sequence and the Cy3-fluorescence was detected with microplate reader. A difference was seen in all of the designed Sox2 -MBs between when the target sequence was present or not. Error bars represent the mean ± SEM. Asterisks denote statistical significance (n = 3 samples, *** p

    Techniques Used: Expressing, Fluorescence, Synthesized, Sequencing

    Detection of Sox2 -MB in undifferentiated mES cells as compared to Sox2 -negative MEFs. (A) Sox2 expression in mES cells and MEFs was analyzed by RT-PCR. Fluorescent signals of (B) MEFs and (C) mES cells treated with Sox2 -MB (blue line) and nonspecific-MB (control, red line) as measured by flow cytometry. Error bars represent the mean ± SEM. Asterisks denotes statistical significance (n = 3 samples, *** p
    Figure Legend Snippet: Detection of Sox2 -MB in undifferentiated mES cells as compared to Sox2 -negative MEFs. (A) Sox2 expression in mES cells and MEFs was analyzed by RT-PCR. Fluorescent signals of (B) MEFs and (C) mES cells treated with Sox2 -MB (blue line) and nonspecific-MB (control, red line) as measured by flow cytometry. Error bars represent the mean ± SEM. Asterisks denotes statistical significance (n = 3 samples, *** p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry, Cytometry

    Detection of Sox2 -MB in differentiated mES cells. (A) mES cells stained for SSEA-1 together with the Sox2 -MB (blue dots) and the nonspecific-MB (red dots). (B) SSEA-1 stained differentiated mES cells treated with Sox2- MB (blue dots) were compared to SSEA1 stained undifferentiated mES treated Sox2- MB (red dots). (C) Undifferentiated mES cells and mES cells differentiated by exposure to RA were analyzed with RT-PCR. (D) Four quadrants (Q1, Q2, Q3 and Q4) of the differentiated mES cells were selected by comparing the nonspecific-MB fluorescent signal with the Sox2 -MB fluorescent signal. (E) The double-positive sorted cell populations (Q2: Sox2-MB + and SSEA1 + ) formed significantly more undifferentiated colonies compared to the positive-negative sorted cell populations (Q1: Sox2-MB - and SSEA1 + Q4: Sox2-MB + and SSEA1 − ), and the double-negative sorted cell population (Q3: Sox2-MB - and SSEA1 − ). (F) Undifferentiated colonies were positively stained for Sox2, Nanog and SSEA1 (Scale bar = 200 µm). Error bars represent the mean ± SEM. Asterisks denotes statistical significance (n = 3 samples ** p
    Figure Legend Snippet: Detection of Sox2 -MB in differentiated mES cells. (A) mES cells stained for SSEA-1 together with the Sox2 -MB (blue dots) and the nonspecific-MB (red dots). (B) SSEA-1 stained differentiated mES cells treated with Sox2- MB (blue dots) were compared to SSEA1 stained undifferentiated mES treated Sox2- MB (red dots). (C) Undifferentiated mES cells and mES cells differentiated by exposure to RA were analyzed with RT-PCR. (D) Four quadrants (Q1, Q2, Q3 and Q4) of the differentiated mES cells were selected by comparing the nonspecific-MB fluorescent signal with the Sox2 -MB fluorescent signal. (E) The double-positive sorted cell populations (Q2: Sox2-MB + and SSEA1 + ) formed significantly more undifferentiated colonies compared to the positive-negative sorted cell populations (Q1: Sox2-MB - and SSEA1 + Q4: Sox2-MB + and SSEA1 − ), and the double-negative sorted cell population (Q3: Sox2-MB - and SSEA1 − ). (F) Undifferentiated colonies were positively stained for Sox2, Nanog and SSEA1 (Scale bar = 200 µm). Error bars represent the mean ± SEM. Asterisks denotes statistical significance (n = 3 samples ** p

    Techniques Used: Staining, Reverse Transcription Polymerase Chain Reaction

    10) Product Images from "The African Zika virus MR-766 is more virulent and causes more severe brain damage than current Asian lineage and dengue virus"

    Article Title: The African Zika virus MR-766 is more virulent and causes more severe brain damage than current Asian lineage and dengue virus

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.156752

    MR-766 infection leads to more cell death in NPCs and neurons than MEX1-44. (A) Confocal imaging of human neural progenitor cells (hNPCs) stained with antibodies against Sox2 (purple) or caspase 3 (green). Scale bars: 20 μm. (B) Quantification of the percentage of caspase 3-positive cells out of total hNPCs. Error bars indicate s.e.m. of results from three independent experiments. Two-way ANOVA revealed a significant difference between mock, MEX1-44 and MR-766 (* P
    Figure Legend Snippet: MR-766 infection leads to more cell death in NPCs and neurons than MEX1-44. (A) Confocal imaging of human neural progenitor cells (hNPCs) stained with antibodies against Sox2 (purple) or caspase 3 (green). Scale bars: 20 μm. (B) Quantification of the percentage of caspase 3-positive cells out of total hNPCs. Error bars indicate s.e.m. of results from three independent experiments. Two-way ANOVA revealed a significant difference between mock, MEX1-44 and MR-766 (* P

    Techniques Used: Infection, Imaging, Staining

    11) Product Images from "Genetic Deletion of Afadin Causes Hydrocephalus by Destruction of Adherens Junctions in Radial Glial and Ependymal Cells in the Midbrain"

    Article Title: Genetic Deletion of Afadin Causes Hydrocephalus by Destruction of Adherens Junctions in Radial Glial and Ependymal Cells in the Midbrain

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0080356

    Distribution of radial glial cells and neurons in the afadin -cKO midbrain. The coronal sections were stained with the anti-Sox2 and anti-class III β-tubulin Abs in the control and afadin -cKO midbrains at E16.5; ( a1 and a2 ) with the anti-Sox2 Ab; and ( b1 and b2 ) with the anti-class III β-tubulin Ab. ( a1–c1 ) The control midbrain; and ( a2–c2 ) the afadin -cKO midbrain. VZ, ventricular zone; IZ, intermediate zone; MZ, mantle zone; Aq, aqueduct. Scale bar: 100 µm.
    Figure Legend Snippet: Distribution of radial glial cells and neurons in the afadin -cKO midbrain. The coronal sections were stained with the anti-Sox2 and anti-class III β-tubulin Abs in the control and afadin -cKO midbrains at E16.5; ( a1 and a2 ) with the anti-Sox2 Ab; and ( b1 and b2 ) with the anti-class III β-tubulin Ab. ( a1–c1 ) The control midbrain; and ( a2–c2 ) the afadin -cKO midbrain. VZ, ventricular zone; IZ, intermediate zone; MZ, mantle zone; Aq, aqueduct. Scale bar: 100 µm.

    Techniques Used: Staining

    12) Product Images from "Cancer Stem-Like Cells in a Case of an Inflammatory Myofibroblastic Tumor of the Lung"

    Article Title: Cancer Stem-Like Cells in a Case of an Inflammatory Myofibroblastic Tumor of the Lung

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2020.00673

    Immunohistochemistry of lung myofibroblastic tumor. Representative immunohistochemical staining of ALDH1A1, SOX2, NANOG, OCT-4, and c-MYC stem cell markers on a myofibroblastic tumor. Images were shown at 10x and 20x magnification.
    Figure Legend Snippet: Immunohistochemistry of lung myofibroblastic tumor. Representative immunohistochemical staining of ALDH1A1, SOX2, NANOG, OCT-4, and c-MYC stem cell markers on a myofibroblastic tumor. Images were shown at 10x and 20x magnification.

    Techniques Used: Immunohistochemistry, Staining

    13) Product Images from "Tranylcypromine Causes Neurotoxicity and Represses BHC110/LSD1 in Human-Induced Pluripotent Stem Cell-Derived Cerebral Organoids Model"

    Article Title: Tranylcypromine Causes Neurotoxicity and Represses BHC110/LSD1 in Human-Induced Pluripotent Stem Cell-Derived Cerebral Organoids Model

    Journal: Frontiers in Neurology

    doi: 10.3389/fneur.2017.00626

    Tranylcypromine treatment impaired neuron and astrocyte of brain organoid. (A,B) Immunostaining for the neural progenitor marker Sox2, astrocyte marker GFAP, and neuron marker TUJ1 in human cerebral organoids after treatment with different concentrations of tranylcypromine. Scale bars: 200 µm.
    Figure Legend Snippet: Tranylcypromine treatment impaired neuron and astrocyte of brain organoid. (A,B) Immunostaining for the neural progenitor marker Sox2, astrocyte marker GFAP, and neuron marker TUJ1 in human cerebral organoids after treatment with different concentrations of tranylcypromine. Scale bars: 200 µm.

    Techniques Used: Immunostaining, Marker

    14) Product Images from "High Efficient Differentiation of Functional Hepatocytes from Porcine Induced Pluripotent Stem Cells"

    Article Title: High Efficient Differentiation of Functional Hepatocytes from Porcine Induced Pluripotent Stem Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0100417

    Generation of piPSCs from PEFs. (A) From left to right, morphology of PEFs, an induced piPSCs colony, and an iPSCs colony post AP staining. Scale bars, 200 µm. (B) Typical iPSC colonies from mouse, pig and human before passaging. Scale bars, 200 µm. (C) Expression of pluripotency markers of Oct4, Nanog, Sox2, SSEA1, SSEA4, TRA 1-60, and TRA 1-81 by immunostaining. Scale bars, 200 µm. (D) Q-PCR analysis of pluripotency marker genes Oct4, Nanog and Sox2 in PEFs (red) and piPSCs (blue). The ratio of ΔΔCT was normalized to the internal control GAPDH, error bars represent SEM of three independent experiments. (E) Histological analysis of teratoma derived from piPSCs. Ectoderm (a): pigment epithelium; Mesoderm (b): muscle; Endoderm (c): ciliated columnar epithelium. (F) Karyotyping analysis show the normal karyotype of piPSCs. Error bars show SEM of three independent experiments. P value was calculated using Student's t -test.
    Figure Legend Snippet: Generation of piPSCs from PEFs. (A) From left to right, morphology of PEFs, an induced piPSCs colony, and an iPSCs colony post AP staining. Scale bars, 200 µm. (B) Typical iPSC colonies from mouse, pig and human before passaging. Scale bars, 200 µm. (C) Expression of pluripotency markers of Oct4, Nanog, Sox2, SSEA1, SSEA4, TRA 1-60, and TRA 1-81 by immunostaining. Scale bars, 200 µm. (D) Q-PCR analysis of pluripotency marker genes Oct4, Nanog and Sox2 in PEFs (red) and piPSCs (blue). The ratio of ΔΔCT was normalized to the internal control GAPDH, error bars represent SEM of three independent experiments. (E) Histological analysis of teratoma derived from piPSCs. Ectoderm (a): pigment epithelium; Mesoderm (b): muscle; Endoderm (c): ciliated columnar epithelium. (F) Karyotyping analysis show the normal karyotype of piPSCs. Error bars show SEM of three independent experiments. P value was calculated using Student's t -test.

    Techniques Used: Staining, Passaging, Expressing, Immunostaining, Polymerase Chain Reaction, Marker, Derivative Assay

    Related Articles

    Immunohistochemistry:

    Article Title: Canal Cristae Growth and Fiber Extension to the Outer Hair Cells of the Mouse Ear Require Prox1 Activity
    Article Snippet: .. Immunohistochemistry Primary antibodies were rabbit anti–β-gal (ICN), rabbit (Covance Research Products) anti–mouse Prox1 (Promega), rat anti–mouse β-tubulin (Sigma), Hoechst nuclear stain (Sigma), Myo VII (gift of T. Hasson, San Diego), Sox2 and BDNF (Invitrogen). .. Secondary antibodies were Alexa 488, 543, and 634–conjugated donkey anti-rabbit (Molecular Probes), Cy3-conjugated donkey anti–guinea pig (Jackson ImmunoResearch Laboratories), and Cy3-conjugated donkey anti-rat (Jackson ImmunoResearch Laboratories) were used predominantly on whole mounted microdissected sensory epithelia .

    Mutagenesis:

    Article Title: Mutations within Sox2/SOX2 are associated with abnormalities in the hypothalamo-pituitary-gonadal axis in mice and humans
    Article Snippet: .. Purified PCR products comprising the entire SOX2 coding region from both affected and unaffected individuals were subcloned downstream of the T7 promoter in the pcDNA3.1(+) vector (Invitrogen) to generate wild-type and mutant proteins for EMSA. .. For transient transfection and cell localization experiments, mutant and wild-type SOX2 was subcloned into the expression vector pCMV/SV-Flag, containing an in-frame N-terminal FLAG epitope.

    Immunohistofluorescence:

    Article Title: Sox2 Is Required for Maintenance and Differentiation of Bronchiolar Clara, Ciliated, and Goblet Cells
    Article Snippet: .. Immunohistofluorescence was performed on 5-µm-thick sections using antibodies generated to Sox2 (1∶200), CCSP (1∶2000, goat polyclonal, sc-9772), acetylated α-tubulin (1∶2000), Ki-67 (1∶100), and secondary antibodies conjugated with Alexa Fluor 594, Alexa Fluor 488 fluorochromes (1∶200, Molecular Probes, Invitrogen Corp., Carlsbad, CA). .. The slides were examined by microscopy using dual fluorescent labeling and a Zeiss Axioplan 2 Imaging Universal Microscope with an Axiocam MRm black and white digital camera (Axiovision Release 4.3).

    Incubation:

    Article Title: Preparation of Induced Pluripotent Stem Cells Using Human Peripheral Blood Monocytes
    Article Snippet: .. To this, anti-human SOX2 (an intranuclear iPS marker protein) rat monoclonal antibody (1:100; Thermo Fisher Scientific, Inc.), anti-human OCT3/4 (an intranuclear iPS marker protein), rabbit polyclonal antibody (1:500; Medical & Biological Laboratories), or anti-human NANOG (an intranuclear iPS marker protein) mouse monoclonal antibody (1:100; Thermo Fisher Scientific, Inc.) was added as primary antibody and incubated at 37°C for 1 hour. .. Alexa Fluor® 594 labeled anti-rat immunoglobulin G (IgG) goat monoclonal antibody, Alexa Fluor 594 labeled anti-mouse IgG donkey monoclonal antibody, or Alexa Fluor 594 labeled anti-rabbit IgG goat monoclonal antibody (1:500; Thermo Fisher Scientific, Inc.) was added as secondary antibody and incubated at 37°C for 1 hour.

    Article Title: SOX2 contributes to melanoma cell invasion
    Article Snippet: .. For immunofluorescence double labeling of xenograft melanomas, 5-μm paraffin sections were incubated with Sox2 and MMP-3 antibodies at 4°C overnight and then incubated with Alexa Fluor 594-conjugated anti-goat IgG and Alexa Fluor 488-conjugated anti-rabbit IgG at room temperature for 1 h. The sections were coverslipped with ProLong Gold anti-fade with DAPI (Invitrogen). .. Sections were analyzed with a BX51/BX52 microscope (Olympus America, Melville, NY, USA), and images were captured using the CytoVision 3.6 software (Applied Imaging, San Jose, CA, USA).

    Article Title: Cancer stem-neuroendocrine cells in an atypical carcinoid case report
    Article Snippet: .. Samples were incubated with the anti-SOX2 (1:200) (MA1-014 Thermo Fisher Scientific, Meridian Road Rockford, IL, USA), overnight at 4 °C. .. Images were collected and the positivity evaluated using a Zeiss Axioskop microscope with a Zeiss AxioCam ICc 3 High-Resolution Microscope Camera.

    Labeling:

    Article Title: SOX2 contributes to melanoma cell invasion
    Article Snippet: .. For immunofluorescence double labeling of xenograft melanomas, 5-μm paraffin sections were incubated with Sox2 and MMP-3 antibodies at 4°C overnight and then incubated with Alexa Fluor 594-conjugated anti-goat IgG and Alexa Fluor 488-conjugated anti-rabbit IgG at room temperature for 1 h. The sections were coverslipped with ProLong Gold anti-fade with DAPI (Invitrogen). .. Sections were analyzed with a BX51/BX52 microscope (Olympus America, Melville, NY, USA), and images were captured using the CytoVision 3.6 software (Applied Imaging, San Jose, CA, USA).

    Purification:

    Article Title: Mutations within Sox2/SOX2 are associated with abnormalities in the hypothalamo-pituitary-gonadal axis in mice and humans
    Article Snippet: .. Purified PCR products comprising the entire SOX2 coding region from both affected and unaffected individuals were subcloned downstream of the T7 promoter in the pcDNA3.1(+) vector (Invitrogen) to generate wild-type and mutant proteins for EMSA. .. For transient transfection and cell localization experiments, mutant and wild-type SOX2 was subcloned into the expression vector pCMV/SV-Flag, containing an in-frame N-terminal FLAG epitope.

    Marker:

    Article Title: Preparation of Induced Pluripotent Stem Cells Using Human Peripheral Blood Monocytes
    Article Snippet: .. To this, anti-human SOX2 (an intranuclear iPS marker protein) rat monoclonal antibody (1:100; Thermo Fisher Scientific, Inc.), anti-human OCT3/4 (an intranuclear iPS marker protein), rabbit polyclonal antibody (1:500; Medical & Biological Laboratories), or anti-human NANOG (an intranuclear iPS marker protein) mouse monoclonal antibody (1:100; Thermo Fisher Scientific, Inc.) was added as primary antibody and incubated at 37°C for 1 hour. .. Alexa Fluor® 594 labeled anti-rat immunoglobulin G (IgG) goat monoclonal antibody, Alexa Fluor 594 labeled anti-mouse IgG donkey monoclonal antibody, or Alexa Fluor 594 labeled anti-rabbit IgG goat monoclonal antibody (1:500; Thermo Fisher Scientific, Inc.) was added as secondary antibody and incubated at 37°C for 1 hour.

    Generated:

    Article Title: Sox2 Is Required for Maintenance and Differentiation of Bronchiolar Clara, Ciliated, and Goblet Cells
    Article Snippet: .. Immunohistofluorescence was performed on 5-µm-thick sections using antibodies generated to Sox2 (1∶200), CCSP (1∶2000, goat polyclonal, sc-9772), acetylated α-tubulin (1∶2000), Ki-67 (1∶100), and secondary antibodies conjugated with Alexa Fluor 594, Alexa Fluor 488 fluorochromes (1∶200, Molecular Probes, Invitrogen Corp., Carlsbad, CA). .. The slides were examined by microscopy using dual fluorescent labeling and a Zeiss Axioplan 2 Imaging Universal Microscope with an Axiocam MRm black and white digital camera (Axiovision Release 4.3).

    Western Blot:

    Article Title: Catalytic-Independent Functions of PARP-1 Determine Sox2 Pioneer Activity at Intractable Genomic Loci
    Article Snippet: .. The eluted immunoprecipitates were run on 8% polyacrylamide-SDS gels, transferred to nitrocellulose, and subjected to Western blotting using antibodies to PARP-1, Sox2, and Oct4 and a chemilumenescent detection system (Thermo Scientific). .. Flag-tagged human PARP-1 proteins (wild-type and DBD point mutant) were expressed in insect cells using a baculovirus expression system and purified by using Flag-affinity purification, as described previously ( ).

    Polymerase Chain Reaction:

    Article Title: Mutations within Sox2/SOX2 are associated with abnormalities in the hypothalamo-pituitary-gonadal axis in mice and humans
    Article Snippet: .. Purified PCR products comprising the entire SOX2 coding region from both affected and unaffected individuals were subcloned downstream of the T7 promoter in the pcDNA3.1(+) vector (Invitrogen) to generate wild-type and mutant proteins for EMSA. .. For transient transfection and cell localization experiments, mutant and wild-type SOX2 was subcloned into the expression vector pCMV/SV-Flag, containing an in-frame N-terminal FLAG epitope.

    Staining:

    Article Title: Canal Cristae Growth and Fiber Extension to the Outer Hair Cells of the Mouse Ear Require Prox1 Activity
    Article Snippet: .. Immunohistochemistry Primary antibodies were rabbit anti–β-gal (ICN), rabbit (Covance Research Products) anti–mouse Prox1 (Promega), rat anti–mouse β-tubulin (Sigma), Hoechst nuclear stain (Sigma), Myo VII (gift of T. Hasson, San Diego), Sox2 and BDNF (Invitrogen). .. Secondary antibodies were Alexa 488, 543, and 634–conjugated donkey anti-rabbit (Molecular Probes), Cy3-conjugated donkey anti–guinea pig (Jackson ImmunoResearch Laboratories), and Cy3-conjugated donkey anti-rat (Jackson ImmunoResearch Laboratories) were used predominantly on whole mounted microdissected sensory epithelia .

    Immunofluorescence:

    Article Title: SOX2 contributes to melanoma cell invasion
    Article Snippet: .. For immunofluorescence double labeling of xenograft melanomas, 5-μm paraffin sections were incubated with Sox2 and MMP-3 antibodies at 4°C overnight and then incubated with Alexa Fluor 594-conjugated anti-goat IgG and Alexa Fluor 488-conjugated anti-rabbit IgG at room temperature for 1 h. The sections were coverslipped with ProLong Gold anti-fade with DAPI (Invitrogen). .. Sections were analyzed with a BX51/BX52 microscope (Olympus America, Melville, NY, USA), and images were captured using the CytoVision 3.6 software (Applied Imaging, San Jose, CA, USA).

    Plasmid Preparation:

    Article Title: Mutations within Sox2/SOX2 are associated with abnormalities in the hypothalamo-pituitary-gonadal axis in mice and humans
    Article Snippet: .. Purified PCR products comprising the entire SOX2 coding region from both affected and unaffected individuals were subcloned downstream of the T7 promoter in the pcDNA3.1(+) vector (Invitrogen) to generate wild-type and mutant proteins for EMSA. .. For transient transfection and cell localization experiments, mutant and wild-type SOX2 was subcloned into the expression vector pCMV/SV-Flag, containing an in-frame N-terminal FLAG epitope.

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  • 95
    Thermo Fisher rabbit anti sox2
    Genome Stability and Pluripotency of Corrected iPSC Lines (A and I) Phase-contrast images of USH2A -USH-iPSC clone B3B1 (A) and USH2A -RP-iPSC clone MS3F7 (I). (B and J) Genomic stability of USH2A -USH-iPSC B3B1 (B) and USH2A -RP-iPSC MS3F7 (J) as determined by a digital qPCR analysis of the most commonly rearranged regions reported in iPSCs. The copy number for each chromosomal position is shown with colored dots. (C–E and K–M) Pluripotency of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the markers OCT3/4 (C and K), <t>SOX2</t> (D and L), and NANOG (E and M), respectively. Scale bars, 50 μM. (F-H and N-P) Differentiation capacity of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the germ layer markers GFAP (ectoderm; F and N), SMA (mesoderm; G and O), and AFP (endoderm; H and P), respectively. Scale bars, 20 μM.
    Rabbit Anti Sox2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti sox2/product/Thermo Fisher
    Average 95 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    rabbit anti sox2 - by Bioz Stars, 2020-07
    95/100 stars
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    93
    Thermo Fisher rat anti sox2
    Western blotting. Protein extractions from 3 LGCA and 3 HGCA EpCAM High (+) and EpCAM Low (-) cell lines were probed for OCT4 (A; 40kDa), <t>SOX2</t> (B; 40-43kDa), NANOG (C; 37-40kDa), KLF4 (D; 54kDa) and c-MYC (E; 42 57kDa). NTERA-2 cells were used as the positive control for all iPSC markers. α-tubulin (F; 50kDa) was used as a loading control. EpCAM High and EpCAM Low cell lines were also probed for their expression of EpCAM (G; bands from ~30-40kDa) and α-SMA (H; 42kDa). HepG2 cells and 3T3 cells were used as the positive control for EpCAM and α-SMA, respectively.
    Rat Anti Sox2, supplied by Thermo Fisher, 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/rat anti sox2/product/Thermo Fisher
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rat anti sox2 - by Bioz Stars, 2020-07
    93/100 stars
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    Image Search Results


    Genome Stability and Pluripotency of Corrected iPSC Lines (A and I) Phase-contrast images of USH2A -USH-iPSC clone B3B1 (A) and USH2A -RP-iPSC clone MS3F7 (I). (B and J) Genomic stability of USH2A -USH-iPSC B3B1 (B) and USH2A -RP-iPSC MS3F7 (J) as determined by a digital qPCR analysis of the most commonly rearranged regions reported in iPSCs. The copy number for each chromosomal position is shown with colored dots. (C–E and K–M) Pluripotency of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the markers OCT3/4 (C and K), SOX2 (D and L), and NANOG (E and M), respectively. Scale bars, 50 μM. (F-H and N-P) Differentiation capacity of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the germ layer markers GFAP (ectoderm; F and N), SMA (mesoderm; G and O), and AFP (endoderm; H and P), respectively. Scale bars, 20 μM.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Genome Editing in Patient iPSCs Corrects the Most Prevalent USH2A Mutations and Reveals Intriguing Mutant mRNA Expression Profiles

    doi: 10.1016/j.omtm.2019.11.016

    Figure Lengend Snippet: Genome Stability and Pluripotency of Corrected iPSC Lines (A and I) Phase-contrast images of USH2A -USH-iPSC clone B3B1 (A) and USH2A -RP-iPSC clone MS3F7 (I). (B and J) Genomic stability of USH2A -USH-iPSC B3B1 (B) and USH2A -RP-iPSC MS3F7 (J) as determined by a digital qPCR analysis of the most commonly rearranged regions reported in iPSCs. The copy number for each chromosomal position is shown with colored dots. (C–E and K–M) Pluripotency of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the markers OCT3/4 (C and K), SOX2 (D and L), and NANOG (E and M), respectively. Scale bars, 50 μM. (F-H and N-P) Differentiation capacity of USH2A -USH-iPSC clone B3B1 and USH2A -RP-iPSC clone MS3F7 as determined by immunostaining of the germ layer markers GFAP (ectoderm; F and N), SMA (mesoderm; G and O), and AFP (endoderm; H and P), respectively. Scale bars, 20 μM.

    Article Snippet: Primary antibodies were used at a 1:200 dilution in blocking solution and incubated overnight at 4°C: rabbit anti-NANOG (Abcam, Paris, France), mouse anti-OCT3/4 (Santa Cruz Biotechnology, Heidelberg, Germany), and rabbit anti-SOX2 (Thermo Fisher Scientific) for the iPSCs, and rabbit anti-GFAP (Dako, Les Ulis, France), mouse anti-SMA (Dako), and mouse anti-AFP (Sigma Aldrich) for the embryoid bodies.

    Techniques: Real-time Polymerase Chain Reaction, Immunostaining

    Expression of Schwann cell markers in peripheral nerves of Sox10 ΔeSC embryos. (A–L) Immunohistochemistry (A–J) and in situ hybridizations (K and L) were performed on transverse sections of wild-type (wt; A, C, E, G, I, and K) and Sox10 ΔeSC (B, D, F, H, J, and L) sciatic nerves at 16.5 (A, B, G, and H) and 18.5 dpc (C–F and I–L) using antibodies directed against Sox2 (A–D), Krox20 (E and F), Oct6 (G–J), and an antisense riboprobe for Mbp (K and L). (C, F, H, and J) Dotted lines indicate the circumference of the sciatic nerve. Bars, 50 µm.

    Journal: The Journal of Cell Biology

    Article Title: Sox10 is required for Schwann cell identity and progression beyond the immature Schwann cell stage

    doi: 10.1083/jcb.200912142

    Figure Lengend Snippet: Expression of Schwann cell markers in peripheral nerves of Sox10 ΔeSC embryos. (A–L) Immunohistochemistry (A–J) and in situ hybridizations (K and L) were performed on transverse sections of wild-type (wt; A, C, E, G, I, and K) and Sox10 ΔeSC (B, D, F, H, J, and L) sciatic nerves at 16.5 (A, B, G, and H) and 18.5 dpc (C–F and I–L) using antibodies directed against Sox2 (A–D), Krox20 (E and F), Oct6 (G–J), and an antisense riboprobe for Mbp (K and L). (C, F, H, and J) Dotted lines indicate the circumference of the sciatic nerve. Bars, 50 µm.

    Article Snippet: For immunohistochemistry, the following primary antibodies were used in various combinations: anti-Sox10 guinea pig antiserum (1:1,000; ), anti-Oct6 rabbit antiserum (1:2,000; ), anti-Krox20 rabbit antiserum (1:200; Covance), anti-Sox2 rabbit antiserum (1:500), anti-Ki67 rabbit antiserum (1:500; Thermo Fisher Scientific), anti-CD3 rabbit antiserum (1:500; Abcam), anti-GFP rabbit antiserum (1:500; Invitrogen), anti-Iba1 rabbit antiserum (1:250; Wako Chemicals USA, Inc.) antidesmin rabbit antiserum (1:1,000; Abcam), anti–von Willebrand factor rabbit antiserum (1:800; Abcam), and anti-PECAM rat antiserum (1:200; BD).

    Techniques: Expressing, Immunohistochemistry, In Situ

    Postnatal development of Sox10 ΔeSC mice and their sciatic nerves. (A) Weight of wild-type (open bars) and age-matched Sox10 ΔeSC (closed bars) mice was monitored at P8, P16, P24, and P32. Data are shown as means ± SEM ( n ≥ 6 for each genotype). Statistically significant differences from wild-type controls were observed from P16 onwards (**, P ≤ 0.01; and ***, P ≤ 0.001 by Student’s t test). (B) Macroscopic appearance of P32 sciatic nerves from wild-type (wt) and Sox10 ΔeSC mice. (C) Sciatic nerve thickness was quantified for wild-type and Sox10 ΔeSC mice at P8, P16, P24, and P32 by determining the area on proximal nerve sections. At least 20 sections from three mice were used per genotype for quantification. Data are presented as mean ± SEM. Differences were statistically significant between wild type and Sox10 ΔeSC mutant from P8 onwards as determined by Student’s t test (**, P ≤ 0.01; ***, P ≤ 0.001). (D–K) Immunohistochemistry was performed on sciatic nerve sections from wild-type (D, F, H, and J) and Sox10 ΔeSC (E, G, I, and K) mice at P16 (D, E, H, and I) and P32 (F, G, J, and K) using antibodies directed against Sox2 (D–G) and Krox20 (H–K). (D, F, I, and K) Dotted lines indicate the circumference of the sciatic nerve. (L–S) In situ hybridization was performed on sciatic nerve sections from wild-type (L, N, P, and R) and Sox10 ΔeSC (M, O, Q, and S) mice at P16 (L, M, P, and Q) and P32 (N, O, R, and S) using antisense riboprobes for Mpz (L–O) and Mbp (P–S). (T–W) Myelin sheaths were visualized by PPD staining of sciatic nerve sections from wild-type (T and V) and Sox10 ΔeSC (U and W) mice at P16 (T and U) and P32 (V and W). (X and Y) Electrophysiology on sciatic nerves of Sox10 ΔeSC mice. Compound action potentials were monopolarly recorded from isolated sciatic nerves of wild-type and Sox10 ΔeSC mice ( n = 2 each). Experiments were performed on both nerves of each animal with identical results within each genotype. Representative superimposed traces are presented for both genotypes showing fast nerve conduction along myelinated fibers (X) and slow conduction along nonmyelinated fibers (Y). The arrows point to components of different conduction velocities (meters/second). Bars: (B) 1 mm; (D–S) 50 µm; (T–W) 3 µm.

    Journal: The Journal of Cell Biology

    Article Title: Sox10 is required for Schwann cell identity and progression beyond the immature Schwann cell stage

    doi: 10.1083/jcb.200912142

    Figure Lengend Snippet: Postnatal development of Sox10 ΔeSC mice and their sciatic nerves. (A) Weight of wild-type (open bars) and age-matched Sox10 ΔeSC (closed bars) mice was monitored at P8, P16, P24, and P32. Data are shown as means ± SEM ( n ≥ 6 for each genotype). Statistically significant differences from wild-type controls were observed from P16 onwards (**, P ≤ 0.01; and ***, P ≤ 0.001 by Student’s t test). (B) Macroscopic appearance of P32 sciatic nerves from wild-type (wt) and Sox10 ΔeSC mice. (C) Sciatic nerve thickness was quantified for wild-type and Sox10 ΔeSC mice at P8, P16, P24, and P32 by determining the area on proximal nerve sections. At least 20 sections from three mice were used per genotype for quantification. Data are presented as mean ± SEM. Differences were statistically significant between wild type and Sox10 ΔeSC mutant from P8 onwards as determined by Student’s t test (**, P ≤ 0.01; ***, P ≤ 0.001). (D–K) Immunohistochemistry was performed on sciatic nerve sections from wild-type (D, F, H, and J) and Sox10 ΔeSC (E, G, I, and K) mice at P16 (D, E, H, and I) and P32 (F, G, J, and K) using antibodies directed against Sox2 (D–G) and Krox20 (H–K). (D, F, I, and K) Dotted lines indicate the circumference of the sciatic nerve. (L–S) In situ hybridization was performed on sciatic nerve sections from wild-type (L, N, P, and R) and Sox10 ΔeSC (M, O, Q, and S) mice at P16 (L, M, P, and Q) and P32 (N, O, R, and S) using antisense riboprobes for Mpz (L–O) and Mbp (P–S). (T–W) Myelin sheaths were visualized by PPD staining of sciatic nerve sections from wild-type (T and V) and Sox10 ΔeSC (U and W) mice at P16 (T and U) and P32 (V and W). (X and Y) Electrophysiology on sciatic nerves of Sox10 ΔeSC mice. Compound action potentials were monopolarly recorded from isolated sciatic nerves of wild-type and Sox10 ΔeSC mice ( n = 2 each). Experiments were performed on both nerves of each animal with identical results within each genotype. Representative superimposed traces are presented for both genotypes showing fast nerve conduction along myelinated fibers (X) and slow conduction along nonmyelinated fibers (Y). The arrows point to components of different conduction velocities (meters/second). Bars: (B) 1 mm; (D–S) 50 µm; (T–W) 3 µm.

    Article Snippet: For immunohistochemistry, the following primary antibodies were used in various combinations: anti-Sox10 guinea pig antiserum (1:1,000; ), anti-Oct6 rabbit antiserum (1:2,000; ), anti-Krox20 rabbit antiserum (1:200; Covance), anti-Sox2 rabbit antiserum (1:500), anti-Ki67 rabbit antiserum (1:500; Thermo Fisher Scientific), anti-CD3 rabbit antiserum (1:500; Abcam), anti-GFP rabbit antiserum (1:500; Invitrogen), anti-Iba1 rabbit antiserum (1:250; Wako Chemicals USA, Inc.) antidesmin rabbit antiserum (1:1,000; Abcam), anti–von Willebrand factor rabbit antiserum (1:800; Abcam), and anti-PECAM rat antiserum (1:200; BD).

    Techniques: Mouse Assay, Mutagenesis, Immunohistochemistry, In Situ Hybridization, Staining, Isolation

    Schwann cell proliferation in sciatic nerves of Sox10 ΔeSC mice. (A and B) The absolute number of Sox2-positive cells was determined in sciatic nerves of Sox10 ΔeSC mice at P8, P16, P24, and P32 (A) and set in relation to the total number of cells (B). (C) Cell numbers were also determined for YFP-expressing cells in sciatic nerves of Dhh::Cre, Rosa26 stopfloxYFP ( Rosa26 DhhYFP ; open bars) and Dhh::Cre, Sox10 fl/fl , Rosa26 stopfloxYFP mice ( Sox10 ΔeSC , Rosa26 DhhYFP ; closed bars) at P8, P16, P24, and P32. (D–G) Coimmunohistochemistry was performed on sciatic nerve sections of Sox10 ΔeSC , Rosa26 DhhYFP mice at P8 (D), P16 (E), P24 (F), and P32 (G) using antibodies against Sox2 (red) and YFP (green). (H) Proliferation rates of wild-type (wt) and Sox10 ΔeSC Schwann cells were determined at P8, P16, P24, and P32 by determining the fraction of Ki67-positive cells among the YFP-labeled cells in Rosa26 DhhYFP (open bars) and Sox10 ΔeSC , Rosa26 DhhYFP mice (closed bars). (I) The proliferation rates of Sox10 ΔeSC Schwann cells (see H) were also used to determine the relative contribution of Schwann cells to the overall proliferation in sciatic nerves of Sox10 ΔeSC mice. For all quantifications, at least 20 sections from three mice were used per genotype. Data are presented as mean ± SEM. According to the Student’s t test, differences were statistically significant between wild type and Sox10 ΔeSC mutant as indicated (**, P ≤ 0.01; ***, P ≤ 0.001). Bars, 10 µm.

    Journal: The Journal of Cell Biology

    Article Title: Sox10 is required for Schwann cell identity and progression beyond the immature Schwann cell stage

    doi: 10.1083/jcb.200912142

    Figure Lengend Snippet: Schwann cell proliferation in sciatic nerves of Sox10 ΔeSC mice. (A and B) The absolute number of Sox2-positive cells was determined in sciatic nerves of Sox10 ΔeSC mice at P8, P16, P24, and P32 (A) and set in relation to the total number of cells (B). (C) Cell numbers were also determined for YFP-expressing cells in sciatic nerves of Dhh::Cre, Rosa26 stopfloxYFP ( Rosa26 DhhYFP ; open bars) and Dhh::Cre, Sox10 fl/fl , Rosa26 stopfloxYFP mice ( Sox10 ΔeSC , Rosa26 DhhYFP ; closed bars) at P8, P16, P24, and P32. (D–G) Coimmunohistochemistry was performed on sciatic nerve sections of Sox10 ΔeSC , Rosa26 DhhYFP mice at P8 (D), P16 (E), P24 (F), and P32 (G) using antibodies against Sox2 (red) and YFP (green). (H) Proliferation rates of wild-type (wt) and Sox10 ΔeSC Schwann cells were determined at P8, P16, P24, and P32 by determining the fraction of Ki67-positive cells among the YFP-labeled cells in Rosa26 DhhYFP (open bars) and Sox10 ΔeSC , Rosa26 DhhYFP mice (closed bars). (I) The proliferation rates of Sox10 ΔeSC Schwann cells (see H) were also used to determine the relative contribution of Schwann cells to the overall proliferation in sciatic nerves of Sox10 ΔeSC mice. For all quantifications, at least 20 sections from three mice were used per genotype. Data are presented as mean ± SEM. According to the Student’s t test, differences were statistically significant between wild type and Sox10 ΔeSC mutant as indicated (**, P ≤ 0.01; ***, P ≤ 0.001). Bars, 10 µm.

    Article Snippet: For immunohistochemistry, the following primary antibodies were used in various combinations: anti-Sox10 guinea pig antiserum (1:1,000; ), anti-Oct6 rabbit antiserum (1:2,000; ), anti-Krox20 rabbit antiserum (1:200; Covance), anti-Sox2 rabbit antiserum (1:500), anti-Ki67 rabbit antiserum (1:500; Thermo Fisher Scientific), anti-CD3 rabbit antiserum (1:500; Abcam), anti-GFP rabbit antiserum (1:500; Invitrogen), anti-Iba1 rabbit antiserum (1:250; Wako Chemicals USA, Inc.) antidesmin rabbit antiserum (1:1,000; Abcam), anti–von Willebrand factor rabbit antiserum (1:800; Abcam), and anti-PECAM rat antiserum (1:200; BD).

    Techniques: Mouse Assay, Expressing, Labeling, Mutagenesis

    Human CLPs directly promote oligodendrogenesis. Human neural stem cells (NSCs) were cultured in the presence of PBS (control), human CHI3L1 and human CHIT1 at 250 ng/ml for 14 days. a – c Cells were immunostained for MBP ( a , oligodendrocytes, green), DCX ( a , neurons, red), NG2 ( b , OPC, green), GFAP ( c , astrocyte, red), SOX2 ( d , NSC, red) and nuclear stain DAPI (blue). Scale bar, 50 μm. e Quantification of MBP + , NG2 + , GFAP + , DCX + , and SOX2 + NSCs after exposure to human Chi3L1or human Chit1. Exposure of differentiating NSCs to CHI3L1 and CHIT1 led to significant increase in oligodendrogenesis. Values are expressed as percent change to PBS-treated NSCs (data are representative of three independent experiments. one-way ANOVA with Dunnett’s multiple comparison test; mean ± s.e.m * p

    Journal: Nature Communications

    Article Title: Chi3l3 induces oligodendrogenesis in an experimental model of autoimmune neuroinflammation

    doi: 10.1038/s41467-018-08140-7

    Figure Lengend Snippet: Human CLPs directly promote oligodendrogenesis. Human neural stem cells (NSCs) were cultured in the presence of PBS (control), human CHI3L1 and human CHIT1 at 250 ng/ml for 14 days. a – c Cells were immunostained for MBP ( a , oligodendrocytes, green), DCX ( a , neurons, red), NG2 ( b , OPC, green), GFAP ( c , astrocyte, red), SOX2 ( d , NSC, red) and nuclear stain DAPI (blue). Scale bar, 50 μm. e Quantification of MBP + , NG2 + , GFAP + , DCX + , and SOX2 + NSCs after exposure to human Chi3L1or human Chit1. Exposure of differentiating NSCs to CHI3L1 and CHIT1 led to significant increase in oligodendrogenesis. Values are expressed as percent change to PBS-treated NSCs (data are representative of three independent experiments. one-way ANOVA with Dunnett’s multiple comparison test; mean ± s.e.m * p

    Article Snippet: For analysis of murine tissue and cells, primary antibodies at working concentrations were: rat anti-BrdU (1:100; Accurate Chemical, cat# YSRTMCA2060GA), rat anti-CD11b (1:50; BD Biosciences, cat# 550282), mouse anti-CD45 (1:100, BioLegend, cat# 103101), rat anti-CD4 (1:200, BD Bioscience, cat# 550278) rabbit anti Chi3l3 (1:50; Stemcell Technology, cat# 60130), rabbit anti-Doublecortin (Dcx, 1:100; Abcam, cat# ab18723), mouse anti-GFAP (1:500; BD Bioscience, cat# 610566), mouse anti-Ki76-FITC (1:100; BD Bioscience, cat# 617472), mouse anti-Map2 (1:250; Sigma-Aldrich, cat# M9942), rabbit anti-NG2 (1:100; Millipore, cat# ab5320), mouse anti-O4 (1:100; Millipore, cat# MAB345), rabbit anti-p-Pyk2 (Tyr402; 1:500, CST, cat# 3291 S), rabbit anti-p-cRaf (Ser259; 1:500, CST, cat# 9421 P), rabbit anti-Sox2 (1:500, Thermo Fisher Scientific, cat# A24339), rabbit anti-p-p38MAPK (THr180/Tyr182; 1:500, CST, cat#4511 P), rabbit anti-p-PLCγ2 (Tyr1217; 1:500, CST, cat# 3871 P), rabbit anti-p-PI(3)K (Tyr458; 1:500, CST cat# 4228 P), rabbit anti-p-Erk1/2 (Thr202/Tyr204; 1:500, CST, cat# 9101), rabbit anti-p-EGFR (Tyr1068; 1:100, CST, cat# 3777).

    Techniques: Cell Culture, Staining

    Chi3l3 directly promotes oligodendrogenesis in vitro. Representative confocal image (above) and quantification (below) of neural stem cells (NSCs) cultured in the presence of PBS (control) or Chi3l3 (100 ng/ml) for 3 days a – d , 5 days e – h , or primary OPCs cultured in the presence of PBS (control) or Chi3l3 (500 ng/ml) for 7 days i . Cells were treated with the nuclear stain TO-PRO-3 (blue) and immunostained for early progenitor markers NG2 ( a ; oligodendrocyte precursor cells, green), GFAP ( b ; astrocytes, green), Dcx ( c ; neuroblasts, green), late progenitor markers O4 ( e ; oligodendrocytes, green), GFAP ( f ; astrocytes, green) and microtubule-associated protein 2 ( g ; Map2, neurons, green), the neural stem cell marker Sox2 ( d , h ; green) and the myelin protein MBP ( i , oligodendrocytes, green). Exposure of differentiating NSCs to Chi3l3 led to significant increase in oligodendrocyte precursor cells and oligodendrocytes, significant decrease in astrocytes, neuroblasts, and neurons and a significant increase in Sox2 + neural stem cells. Scale bar, 50 μm. Inserts show representative cells. Scale bar, 20 μm. j – n Gene expression of Cspg4 (NG2; j ), Gfap k , and Map2 ( l ; 3 days) and Ccnd1 and Ccnd2 ( m , n ; 24 h) mRNA in PBS (control) or Chi3l3-treated differentiating NSCs. Values were normalized against Gapdh (AU, arbitrary unit; n.s., not significant;). Number o and size p of neurospheres from NSCs exposed to Chi3l3 or PBS (control). (n.s., not significant; two-tailed Student’s t test; data are representative of three independent experiments with n = 5 replicates a , c , d , i , n = 9 (Control) and 10 (Chi3l3) replicates b , n = 12 (Control) and 9 (Chi3l3) replicates e , n = 12 replicates f , g , n = 3 replicates h , j , l , m , n ), n = 3 (Control) and 2 (Chi3l3) replicates k , n = 4 replicates o , n = 8 (control) and four (Chi3l3) replicates p ). mean ± s.e.m. * p

    Journal: Nature Communications

    Article Title: Chi3l3 induces oligodendrogenesis in an experimental model of autoimmune neuroinflammation

    doi: 10.1038/s41467-018-08140-7

    Figure Lengend Snippet: Chi3l3 directly promotes oligodendrogenesis in vitro. Representative confocal image (above) and quantification (below) of neural stem cells (NSCs) cultured in the presence of PBS (control) or Chi3l3 (100 ng/ml) for 3 days a – d , 5 days e – h , or primary OPCs cultured in the presence of PBS (control) or Chi3l3 (500 ng/ml) for 7 days i . Cells were treated with the nuclear stain TO-PRO-3 (blue) and immunostained for early progenitor markers NG2 ( a ; oligodendrocyte precursor cells, green), GFAP ( b ; astrocytes, green), Dcx ( c ; neuroblasts, green), late progenitor markers O4 ( e ; oligodendrocytes, green), GFAP ( f ; astrocytes, green) and microtubule-associated protein 2 ( g ; Map2, neurons, green), the neural stem cell marker Sox2 ( d , h ; green) and the myelin protein MBP ( i , oligodendrocytes, green). Exposure of differentiating NSCs to Chi3l3 led to significant increase in oligodendrocyte precursor cells and oligodendrocytes, significant decrease in astrocytes, neuroblasts, and neurons and a significant increase in Sox2 + neural stem cells. Scale bar, 50 μm. Inserts show representative cells. Scale bar, 20 μm. j – n Gene expression of Cspg4 (NG2; j ), Gfap k , and Map2 ( l ; 3 days) and Ccnd1 and Ccnd2 ( m , n ; 24 h) mRNA in PBS (control) or Chi3l3-treated differentiating NSCs. Values were normalized against Gapdh (AU, arbitrary unit; n.s., not significant;). Number o and size p of neurospheres from NSCs exposed to Chi3l3 or PBS (control). (n.s., not significant; two-tailed Student’s t test; data are representative of three independent experiments with n = 5 replicates a , c , d , i , n = 9 (Control) and 10 (Chi3l3) replicates b , n = 12 (Control) and 9 (Chi3l3) replicates e , n = 12 replicates f , g , n = 3 replicates h , j , l , m , n ), n = 3 (Control) and 2 (Chi3l3) replicates k , n = 4 replicates o , n = 8 (control) and four (Chi3l3) replicates p ). mean ± s.e.m. * p

    Article Snippet: For analysis of murine tissue and cells, primary antibodies at working concentrations were: rat anti-BrdU (1:100; Accurate Chemical, cat# YSRTMCA2060GA), rat anti-CD11b (1:50; BD Biosciences, cat# 550282), mouse anti-CD45 (1:100, BioLegend, cat# 103101), rat anti-CD4 (1:200, BD Bioscience, cat# 550278) rabbit anti Chi3l3 (1:50; Stemcell Technology, cat# 60130), rabbit anti-Doublecortin (Dcx, 1:100; Abcam, cat# ab18723), mouse anti-GFAP (1:500; BD Bioscience, cat# 610566), mouse anti-Ki76-FITC (1:100; BD Bioscience, cat# 617472), mouse anti-Map2 (1:250; Sigma-Aldrich, cat# M9942), rabbit anti-NG2 (1:100; Millipore, cat# ab5320), mouse anti-O4 (1:100; Millipore, cat# MAB345), rabbit anti-p-Pyk2 (Tyr402; 1:500, CST, cat# 3291 S), rabbit anti-p-cRaf (Ser259; 1:500, CST, cat# 9421 P), rabbit anti-Sox2 (1:500, Thermo Fisher Scientific, cat# A24339), rabbit anti-p-p38MAPK (THr180/Tyr182; 1:500, CST, cat#4511 P), rabbit anti-p-PLCγ2 (Tyr1217; 1:500, CST, cat# 3871 P), rabbit anti-p-PI(3)K (Tyr458; 1:500, CST cat# 4228 P), rabbit anti-p-Erk1/2 (Thr202/Tyr204; 1:500, CST, cat# 9101), rabbit anti-p-EGFR (Tyr1068; 1:100, CST, cat# 3777).

    Techniques: In Vitro, Cell Culture, Staining, Marker, Expressing, Two Tailed Test

    Western blotting. Protein extractions from 3 LGCA and 3 HGCA EpCAM High (+) and EpCAM Low (-) cell lines were probed for OCT4 (A; 40kDa), SOX2 (B; 40-43kDa), NANOG (C; 37-40kDa), KLF4 (D; 54kDa) and c-MYC (E; 42 57kDa). NTERA-2 cells were used as the positive control for all iPSC markers. α-tubulin (F; 50kDa) was used as a loading control. EpCAM High and EpCAM Low cell lines were also probed for their expression of EpCAM (G; bands from ~30-40kDa) and α-SMA (H; 42kDa). HepG2 cells and 3T3 cells were used as the positive control for EpCAM and α-SMA, respectively.

    Journal: PLoS ONE

    Article Title: Colon adenocarcinoma-derived cells that express induced-pluripotent stem cell markers possess stem cell function

    doi: 10.1371/journal.pone.0232934

    Figure Lengend Snippet: Western blotting. Protein extractions from 3 LGCA and 3 HGCA EpCAM High (+) and EpCAM Low (-) cell lines were probed for OCT4 (A; 40kDa), SOX2 (B; 40-43kDa), NANOG (C; 37-40kDa), KLF4 (D; 54kDa) and c-MYC (E; 42 57kDa). NTERA-2 cells were used as the positive control for all iPSC markers. α-tubulin (F; 50kDa) was used as a loading control. EpCAM High and EpCAM Low cell lines were also probed for their expression of EpCAM (G; bands from ~30-40kDa) and α-SMA (H; 42kDa). HepG2 cells and 3T3 cells were used as the positive control for EpCAM and α-SMA, respectively.

    Article Snippet: Primary antibodies included rabbit anti-OCT4 (cat# A24867, Thermo), rat anti-SOX2 (cat# A24759, Thermo), mouse IgG3 anti-SSEA4 (cat# A24866, Thermo) and mouse IgM anti-TRA-1-60 (cat# A24868, Thermo).

    Techniques: Western Blot, Positive Control, Expressing

    RT-qPCR. RNA was extracted from EpCAM High and EpCAM Low cells lines from 3 LGCA and 3 HGCA cases, and RT-qPCR was carried out to measure the mRNA levels of OCT4 (A), SOX2 (B), NANOG (C), KLF4 (D) and c-MYC (E). Triplicate values are shown by dots (EpCAM Low ) and squares (EpCAM High ), with mean and 95% confidence intervals. Abundance was measured relative to qPCR Human Reference Total RNA (Mediray). LGCA (n = 3); HGCA (n = 3).

    Journal: PLoS ONE

    Article Title: Colon adenocarcinoma-derived cells that express induced-pluripotent stem cell markers possess stem cell function

    doi: 10.1371/journal.pone.0232934

    Figure Lengend Snippet: RT-qPCR. RNA was extracted from EpCAM High and EpCAM Low cells lines from 3 LGCA and 3 HGCA cases, and RT-qPCR was carried out to measure the mRNA levels of OCT4 (A), SOX2 (B), NANOG (C), KLF4 (D) and c-MYC (E). Triplicate values are shown by dots (EpCAM Low ) and squares (EpCAM High ), with mean and 95% confidence intervals. Abundance was measured relative to qPCR Human Reference Total RNA (Mediray). LGCA (n = 3); HGCA (n = 3).

    Article Snippet: Primary antibodies included rabbit anti-OCT4 (cat# A24867, Thermo), rat anti-SOX2 (cat# A24759, Thermo), mouse IgG3 anti-SSEA4 (cat# A24866, Thermo) and mouse IgM anti-TRA-1-60 (cat# A24868, Thermo).

    Techniques: Quantitative RT-PCR, Real-time Polymerase Chain Reaction

    Densitometry performed on western blot. Densitometry data provided semi-quantitative data for protein abundance. The intensity values of all LGCA cell lines (both EpCAM High and EpCAM Low ) and all HGCA cell lines (both EpCAM High and EpCAM Low ) were combined and the average intensity calculated, and these are shown for OCT4 (A), SOX2 (C), NANOG (E), KLF4 (G) and c-MYC (I). The intensity values of all EpCAM Low cell lines (both LGCA and HGCA) and all EpCAM High cell lines (both LGCA and HGCA) were combined and the average intensity calculated, and these are shown for OCT4 (B), SOX2 (D), NANOG (F), KLF4 (H), c-MYC (J), EpCAM (K) and α-SMA (L). Individual intensity values were normalized against the loading control α-tubulin before being combined and averaged. Error bars show standard deviation.

    Journal: PLoS ONE

    Article Title: Colon adenocarcinoma-derived cells that express induced-pluripotent stem cell markers possess stem cell function

    doi: 10.1371/journal.pone.0232934

    Figure Lengend Snippet: Densitometry performed on western blot. Densitometry data provided semi-quantitative data for protein abundance. The intensity values of all LGCA cell lines (both EpCAM High and EpCAM Low ) and all HGCA cell lines (both EpCAM High and EpCAM Low ) were combined and the average intensity calculated, and these are shown for OCT4 (A), SOX2 (C), NANOG (E), KLF4 (G) and c-MYC (I). The intensity values of all EpCAM Low cell lines (both LGCA and HGCA) and all EpCAM High cell lines (both LGCA and HGCA) were combined and the average intensity calculated, and these are shown for OCT4 (B), SOX2 (D), NANOG (F), KLF4 (H), c-MYC (J), EpCAM (K) and α-SMA (L). Individual intensity values were normalized against the loading control α-tubulin before being combined and averaged. Error bars show standard deviation.

    Article Snippet: Primary antibodies included rabbit anti-OCT4 (cat# A24867, Thermo), rat anti-SOX2 (cat# A24759, Thermo), mouse IgG3 anti-SSEA4 (cat# A24866, Thermo) and mouse IgM anti-TRA-1-60 (cat# A24868, Thermo).

    Techniques: Western Blot, Standard Deviation

    Representative immunofluorescence immunocytochemical images. EpCAM Low (A) and EpCAM High (B) cells from low-grade colon adenocarcinoma (LGCA)-derived primary cell lines, and EpCAM Low (C) and EpCAM High (D) cells from high-grade colon adenocarcinoma (HGCA)-derived primary cell lines showing expression of SOX2 (green) and TRA-1-60 (red). LGCA (n = 3); HGCA (n = 3). Positive control NTERA-2 (E) and CaCo2 (F) cells were stained for SOX2 (green) and TRA-1-60 (red) (B). Original magnification: 400x; scale bar = 20 μm.

    Journal: PLoS ONE

    Article Title: Colon adenocarcinoma-derived cells that express induced-pluripotent stem cell markers possess stem cell function

    doi: 10.1371/journal.pone.0232934

    Figure Lengend Snippet: Representative immunofluorescence immunocytochemical images. EpCAM Low (A) and EpCAM High (B) cells from low-grade colon adenocarcinoma (LGCA)-derived primary cell lines, and EpCAM Low (C) and EpCAM High (D) cells from high-grade colon adenocarcinoma (HGCA)-derived primary cell lines showing expression of SOX2 (green) and TRA-1-60 (red). LGCA (n = 3); HGCA (n = 3). Positive control NTERA-2 (E) and CaCo2 (F) cells were stained for SOX2 (green) and TRA-1-60 (red) (B). Original magnification: 400x; scale bar = 20 μm.

    Article Snippet: Primary antibodies included rabbit anti-OCT4 (cat# A24867, Thermo), rat anti-SOX2 (cat# A24759, Thermo), mouse IgG3 anti-SSEA4 (cat# A24866, Thermo) and mouse IgM anti-TRA-1-60 (cat# A24868, Thermo).

    Techniques: Immunofluorescence, Derivative Assay, Expressing, Positive Control, Staining