anti sox2  (Abcam)

 
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    Name:
    Anti SOX2 antibody EPR3131
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    ab92494
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    Structured Review

    Abcam anti sox2
    Spatial analysis of the primitive streak region. (A) Unique DEGs to each E8.5 region (≥1.5-fold change compared with other samples; for unique DEGs in St5, see Table S1). Asterisk indicates that no specific in situ probe could be constructed. (B) Bhmt2 and Ccno expression in the ventral (V) but not dorsal (D) NSB layer. (C) Whole-mount in situ hybridisation for Mnx1 . tg, tail gut; vn, ventral node. (D) <t>Sox2</t> versus T intensity values for all samples. Abbreviations are preceded by embryo age; colours indicates fate. Regions of similar fate are enclosed by a coloured line.

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    Images

    1) Product Images from "Transcriptionally dynamic progenitor populations organised around a stable niche drive axial patterning"

    Article Title: Transcriptionally dynamic progenitor populations organised around a stable niche drive axial patterning

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.168161

    Spatial analysis of the primitive streak region. (A) Unique DEGs to each E8.5 region (≥1.5-fold change compared with other samples; for unique DEGs in St5, see Table S1). Asterisk indicates that no specific in situ probe could be constructed. (B) Bhmt2 and Ccno expression in the ventral (V) but not dorsal (D) NSB layer. (C) Whole-mount in situ hybridisation for Mnx1 . tg, tail gut; vn, ventral node. (D) Sox2 versus T intensity values for all samples. Abbreviations are preceded by embryo age; colours indicates fate. Regions of similar fate are enclosed by a coloured line.
    Figure Legend Snippet: Spatial analysis of the primitive streak region. (A) Unique DEGs to each E8.5 region (≥1.5-fold change compared with other samples; for unique DEGs in St5, see Table S1). Asterisk indicates that no specific in situ probe could be constructed. (B) Bhmt2 and Ccno expression in the ventral (V) but not dorsal (D) NSB layer. (C) Whole-mount in situ hybridisation for Mnx1 . tg, tail gut; vn, ventral node. (D) Sox2 versus T intensity values for all samples. Abbreviations are preceded by embryo age; colours indicates fate. Regions of similar fate are enclosed by a coloured line.

    Techniques Used: In Situ, Construct, Expressing, Hybridization

    NotoPs are essential for correct axis elongation. (A) DiI labelling of the ventral node or crown at E8.5 (2-5 s; Aa,Ae). (Aa′,Ae′) Magnified view of the ventral node region. (Aa″,Ae″) Red channel showing DiI labelling. The same embryos are shown after 48 h (Ab) or 24 h (Af) ex vivo culture. Ab′ and Af' show magnified views of Ab and Af respectively. (Af″) Red channel showing DiI labelling. DiI was found in the notochord and dorsal gut (Ac,Ad,Ag,Ah). Insets in Ac, Ad, Ag and Ah show magnified views of DiI labelling in notochord and gut. (Ai) DiI labelling per embryo after culture with average anterior limit (±s.d.). The anterior limit in 4/6 crown-labelled embryos was in the presomitic mesoderm. The anterior limit in each of these embryos is denoted as (total somites/embryo) +1. As the presomitic mesoderm contains ∼7 presomites, the average anterior limit may be posterior to s17. Noto, notochord; NT, neural tube; PXM, paraxial mesoderm. Asterisk indicates 2/6 embryos had a minor contribution in the posterior neural tube. (B) Sox2/T immunostained section of embryo labelled with DiI in the crown after 24 h in culture (n embryos ). (D) Sagittal confocal sections through CLE-electroporated (Da), and NSB-electroporated class I (Db) and II (Dc) embryos after 24 h, with the notochord (noto) end shown in yellow. nt, neural tube; som, somite.
    Figure Legend Snippet: NotoPs are essential for correct axis elongation. (A) DiI labelling of the ventral node or crown at E8.5 (2-5 s; Aa,Ae). (Aa′,Ae′) Magnified view of the ventral node region. (Aa″,Ae″) Red channel showing DiI labelling. The same embryos are shown after 48 h (Ab) or 24 h (Af) ex vivo culture. Ab′ and Af' show magnified views of Ab and Af respectively. (Af″) Red channel showing DiI labelling. DiI was found in the notochord and dorsal gut (Ac,Ad,Ag,Ah). Insets in Ac, Ad, Ag and Ah show magnified views of DiI labelling in notochord and gut. (Ai) DiI labelling per embryo after culture with average anterior limit (±s.d.). The anterior limit in 4/6 crown-labelled embryos was in the presomitic mesoderm. The anterior limit in each of these embryos is denoted as (total somites/embryo) +1. As the presomitic mesoderm contains ∼7 presomites, the average anterior limit may be posterior to s17. Noto, notochord; NT, neural tube; PXM, paraxial mesoderm. Asterisk indicates 2/6 embryos had a minor contribution in the posterior neural tube. (B) Sox2/T immunostained section of embryo labelled with DiI in the crown after 24 h in culture (n embryos ). (D) Sagittal confocal sections through CLE-electroporated (Da), and NSB-electroporated class I (Db) and II (Dc) embryos after 24 h, with the notochord (noto) end shown in yellow. nt, neural tube; som, somite.

    Techniques Used: Ex Vivo

    2) Product Images from "Neuroprotective Properties of Asiatic Acid against 5-Fluorouracil Chemotherapy in the Hippocampus in an Adult Rat Model"

    Article Title: Neuroprotective Properties of Asiatic Acid against 5-Fluorouracil Chemotherapy in the Hippocampus in an Adult Rat Model

    Journal: Nutrients

    doi: 10.3390/nu10081053

    Mean ± SEM of Notch1 ( A ), sex determining region Y-box 2 or SOX2 ( B ), nestin ( C ) and doublecortin or DCX ( D ) protein expressions in the hippocampus were assessed by immunoblotting. * p
    Figure Legend Snippet: Mean ± SEM of Notch1 ( A ), sex determining region Y-box 2 or SOX2 ( B ), nestin ( C ) and doublecortin or DCX ( D ) protein expressions in the hippocampus were assessed by immunoblotting. * p

    Techniques Used:

    3) Product Images from "Maturation, not initiation, is the major roadblock during reprogramming toward pluripotency from human fibroblasts"

    Article Title: Maturation, not initiation, is the major roadblock during reprogramming toward pluripotency from human fibroblasts

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1310291110

    Model of the reprogramming process. Black dots, HDFs or nontransduced cells; green dots, transduced cells with SOX2-IRES-EGFP; magenta dots, TRA-1-60 (+) cells.
    Figure Legend Snippet: Model of the reprogramming process. Black dots, HDFs or nontransduced cells; green dots, transduced cells with SOX2-IRES-EGFP; magenta dots, TRA-1-60 (+) cells.

    Techniques Used:

    Efficiency of iPSC induction. ( A ) Experimental scheme to analyze the reprogramming process. Magenta dots, TRA-1-60 (+) cells; green dots, transduced cells with SOX2-IRES-EGFP; black dots, HDFs or nontransduced cells. HDFs were introduced with OKM plus SOX2-IRES-EGFP. Transduced cells were replated at day 7 on feeder cells to analyze the generating efficiency of iPSC colonies on day 24. TRA-1-60 (+) cells, EGFP(+)/TRA-1-60 (−) cells, and EGFP(−)/TRA-1-60 (−) cells were sorted using MACS and FACS on days 7, 11, and 15. Sorted TRA-1-60 (+) cells on day 11 were replated on feeder cells to analyze the TRA-1-60 (+) cells on days 20 and 28 and (−) cells on day 20. ( B ) Proportion of EGFP (+) cells 7 d after OSKM transduction into various HDF lines was analyzed by flow cytometry. The HDFs were derived from various ages (y, year; m, month) of Caucasian and Japanese males (M) and females (F). n = 3. Error bars indicate SD. ( C ) Number of integrated retroviral transgenes per TRA-1-60 (+), EGFP (+)/TRA-1-60 (−), and EGFP (−)/TRA-1-60 (−) cell on days 7, 11, and 15 was analyzed by a quantitative genomic PCR analysis. Parental nontransduced HDFs were used as a negative control. n = 3. Error bars indicate SD. Numbers of integration in seven established iPSC lines were averaged. ( D ) Protein levels of OCT3/4, SOX2, KLF4, and c-MYC in TRA-1-60 (+), EGFP (+)/TRA-1-60 (−), and EGFP (−)/TRA-1-60 (−) cells on day 7. ( E ) Quantification of protein levels in D . Protein levels were adjusted by β-actin for quantification. Protein levels of OCT3/4, SOX2, and c-MYC were normalized with those in HDFs. Protein levels of KLF4 from upper (magenta) and lower (blue) bands are shown separately. Protein levels of KLF4 were normalized with that of upper band in HDFs. n = 3. Error bars indicate SD. ( F ) Relative RNA expression of transgenous (green) and endogenous (yellow) KLF4 on day 7. All values were normalized with total expression of KLF4 in HDFs. n = 3. Error bars indicate SD. ( G ) Number of iPSC colonies derived from 2.5 × 10 5 HDFs transduced with OSKM on day 24. The HDFs were derived from various ages (y, year; m, month) of Caucasian and Japanese males (M) and females (F). n = 3. Error bars indicate SD.
    Figure Legend Snippet: Efficiency of iPSC induction. ( A ) Experimental scheme to analyze the reprogramming process. Magenta dots, TRA-1-60 (+) cells; green dots, transduced cells with SOX2-IRES-EGFP; black dots, HDFs or nontransduced cells. HDFs were introduced with OKM plus SOX2-IRES-EGFP. Transduced cells were replated at day 7 on feeder cells to analyze the generating efficiency of iPSC colonies on day 24. TRA-1-60 (+) cells, EGFP(+)/TRA-1-60 (−) cells, and EGFP(−)/TRA-1-60 (−) cells were sorted using MACS and FACS on days 7, 11, and 15. Sorted TRA-1-60 (+) cells on day 11 were replated on feeder cells to analyze the TRA-1-60 (+) cells on days 20 and 28 and (−) cells on day 20. ( B ) Proportion of EGFP (+) cells 7 d after OSKM transduction into various HDF lines was analyzed by flow cytometry. The HDFs were derived from various ages (y, year; m, month) of Caucasian and Japanese males (M) and females (F). n = 3. Error bars indicate SD. ( C ) Number of integrated retroviral transgenes per TRA-1-60 (+), EGFP (+)/TRA-1-60 (−), and EGFP (−)/TRA-1-60 (−) cell on days 7, 11, and 15 was analyzed by a quantitative genomic PCR analysis. Parental nontransduced HDFs were used as a negative control. n = 3. Error bars indicate SD. Numbers of integration in seven established iPSC lines were averaged. ( D ) Protein levels of OCT3/4, SOX2, KLF4, and c-MYC in TRA-1-60 (+), EGFP (+)/TRA-1-60 (−), and EGFP (−)/TRA-1-60 (−) cells on day 7. ( E ) Quantification of protein levels in D . Protein levels were adjusted by β-actin for quantification. Protein levels of OCT3/4, SOX2, and c-MYC were normalized with those in HDFs. Protein levels of KLF4 from upper (magenta) and lower (blue) bands are shown separately. Protein levels of KLF4 were normalized with that of upper band in HDFs. n = 3. Error bars indicate SD. ( F ) Relative RNA expression of transgenous (green) and endogenous (yellow) KLF4 on day 7. All values were normalized with total expression of KLF4 in HDFs. n = 3. Error bars indicate SD. ( G ) Number of iPSC colonies derived from 2.5 × 10 5 HDFs transduced with OSKM on day 24. The HDFs were derived from various ages (y, year; m, month) of Caucasian and Japanese males (M) and females (F). n = 3. Error bars indicate SD.

    Techniques Used: Magnetic Cell Separation, FACS, Transduction, Flow Cytometry, Cytometry, Derivative Assay, Polymerase Chain Reaction, Negative Control, RNA Expression, Expressing

    4) Product Images from "Chd2 regulates chromatin for proper gene expression toward differentiation in mouse embryonic stem cells"

    Article Title: Chd2 regulates chromatin for proper gene expression toward differentiation in mouse embryonic stem cells

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkx475

    Chd2 depletion is independent of the capacity for self-renewal in undifferentiated mESCs. ( A ) Scheme of generating Chd2 mut/mut ESCs using the CRISPR/Cas9 system ( 25 ). The gRNA target site (chr. 7: 80, 664, 568–80, 664, 590) is indicated by a pink box. Details are shown in Supplementary Figure S1A . ( B ) Depletion of endogenous Chd2 protein in ESCs using the CRISPR/Cas9 system. Western blot analyses of WT and Chd2 mut/mut ESCs using antibodies against Chd2, Brg1, Oct3/4, Nanog, Sox2 and Hsp90 are shown. ( C ) Morphologies of WT and Chd2 mut/mut ESCs were not significantly changed under a bright field. Scale bars = 50 μm. ( D ) Cell cycle analysis of WT and Chd2 mut/mut ESCs by flow cytometry. The population of cells in each cell cycle phase (G 1 , S or G 2 /M) was calculated based on the intensity of Hoechst 33342. Data represent the mean of three independent experiments ± standard deviation. n.s., P > 0.05. ( E ) Scatter plot showing the expression levels of all genes in undifferentiated WT and Chd2 mut/mut ESCs. RNA-seq datasets were obtained and FPKM values were calculated. The red line indicates a regression line. ( F ) Venn diagram showing the overlap between genes with FPKM > 5 in WT and Chd2 mut/mut ESCs.
    Figure Legend Snippet: Chd2 depletion is independent of the capacity for self-renewal in undifferentiated mESCs. ( A ) Scheme of generating Chd2 mut/mut ESCs using the CRISPR/Cas9 system ( 25 ). The gRNA target site (chr. 7: 80, 664, 568–80, 664, 590) is indicated by a pink box. Details are shown in Supplementary Figure S1A . ( B ) Depletion of endogenous Chd2 protein in ESCs using the CRISPR/Cas9 system. Western blot analyses of WT and Chd2 mut/mut ESCs using antibodies against Chd2, Brg1, Oct3/4, Nanog, Sox2 and Hsp90 are shown. ( C ) Morphologies of WT and Chd2 mut/mut ESCs were not significantly changed under a bright field. Scale bars = 50 μm. ( D ) Cell cycle analysis of WT and Chd2 mut/mut ESCs by flow cytometry. The population of cells in each cell cycle phase (G 1 , S or G 2 /M) was calculated based on the intensity of Hoechst 33342. Data represent the mean of three independent experiments ± standard deviation. n.s., P > 0.05. ( E ) Scatter plot showing the expression levels of all genes in undifferentiated WT and Chd2 mut/mut ESCs. RNA-seq datasets were obtained and FPKM values were calculated. The red line indicates a regression line. ( F ) Venn diagram showing the overlap between genes with FPKM > 5 in WT and Chd2 mut/mut ESCs.

    Techniques Used: CRISPR, Western Blot, Cell Cycle Assay, Flow Cytometry, Cytometry, Standard Deviation, Expressing, RNA Sequencing Assay

    Chd2 is associated with Oct3/4 for H3.3 deposition in developmental gene loci. ( A ) Immunoprecipitations were performed from WT ESC extracts using a Chd2-specific antibody or IgG as a control. Western blot analyses of the products using antibodies against Chd2, Oct3/4, Sox2 and H3.3 are shown. ( B ) Motif analysis of the Chd2-binding region around the bivalent genes (±50 kbp) using HOMER ( 43 ). ( C ) ChIP-qPCR assay using anti-Chd2 antibodies at bivalent genes ( Gata6, T, Nes, Nanog and Myh1 ) in WT and Oct3/4-depleted mESCs. Recovery efficiency (mean ± standard deviation of three independent experiments) is expressed as enrichment relative to the input. * P
    Figure Legend Snippet: Chd2 is associated with Oct3/4 for H3.3 deposition in developmental gene loci. ( A ) Immunoprecipitations were performed from WT ESC extracts using a Chd2-specific antibody or IgG as a control. Western blot analyses of the products using antibodies against Chd2, Oct3/4, Sox2 and H3.3 are shown. ( B ) Motif analysis of the Chd2-binding region around the bivalent genes (±50 kbp) using HOMER ( 43 ). ( C ) ChIP-qPCR assay using anti-Chd2 antibodies at bivalent genes ( Gata6, T, Nes, Nanog and Myh1 ) in WT and Oct3/4-depleted mESCs. Recovery efficiency (mean ± standard deviation of three independent experiments) is expressed as enrichment relative to the input. * P

    Techniques Used: Western Blot, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Standard Deviation

    5) Product Images from "AFF4 promotes tumorigenesis and tumor-initiation capacity of head and neck squamous cell carcinoma cells by regulating SOX2"

    Article Title: AFF4 promotes tumorigenesis and tumor-initiation capacity of head and neck squamous cell carcinoma cells by regulating SOX2

    Journal: Carcinogenesis

    doi: 10.1093/carcin/bgy046

    AFF4 promotes SOX2 expression and recruits CDK9 to SOX2 promoter. ( a ) Western blot of SOX2 expression in response to depletion of AFF4 at 48 h post transfection. n = 3. ( b ) Western blot of SOX2 expression in response to overexpression of AFF4 at 48 h post transfection. n = 3. ( c , d ) ChIP assay was conducted to study the recruitment of AFF4 at SOX2 promoter in SCC1 cells (c) and SCC23 cells (d) transfected with SCR and siAFF4 at 48 h post transfection. n = 3. ( e , f ) ChIP assay was conducted to study the recruitment of CDK9 at SOX2 promoter in SCC1 cells (e) and SCC23 cells (f) transfected with SCR and siAFF4 at 48 h post transfection. n = 3. ( g , h ) ChIP assay was conducted to study the H3K4me3 levels at SOX2 promoter in SCC1 cells (g) and SCC23 cells (h) transfected with SCR and siAFF4 at 48 h post transfection, respectively. n = 3. * P -value
    Figure Legend Snippet: AFF4 promotes SOX2 expression and recruits CDK9 to SOX2 promoter. ( a ) Western blot of SOX2 expression in response to depletion of AFF4 at 48 h post transfection. n = 3. ( b ) Western blot of SOX2 expression in response to overexpression of AFF4 at 48 h post transfection. n = 3. ( c , d ) ChIP assay was conducted to study the recruitment of AFF4 at SOX2 promoter in SCC1 cells (c) and SCC23 cells (d) transfected with SCR and siAFF4 at 48 h post transfection. n = 3. ( e , f ) ChIP assay was conducted to study the recruitment of CDK9 at SOX2 promoter in SCC1 cells (e) and SCC23 cells (f) transfected with SCR and siAFF4 at 48 h post transfection. n = 3. ( g , h ) ChIP assay was conducted to study the H3K4me3 levels at SOX2 promoter in SCC1 cells (g) and SCC23 cells (h) transfected with SCR and siAFF4 at 48 h post transfection, respectively. n = 3. * P -value

    Techniques Used: Expressing, Western Blot, Transfection, Over Expression, Chromatin Immunoprecipitation

    AFF4 enhances tumor-initiation capacity of HNSCC cells. ( a , b ) Flow cytometry assay performed to examine the effects of AFF4 depletion on ALDH activity of SCC1 cells (a) and SCC23 cells (b). Cells stained with ALDH+DEAB, an ALDH inhibitor, served as the negative control. ( c ) Quantification of ALDH + cells in a and b. n = 3. ( d ) Sphere formation assay conducted to examine the effects of AFF4 depletion on sphere formation capacity of SCC1 cells and SCC23 cells. Bar indicates 200 μm. ( e ) Quantification of formed spheres (diameter exceeding 10 µm) in d. n = 3. ( f ) qPCR analysis of the effects of AFF4 depletion on expression of BMI1 , CD44 , NANOG and SOX2 in SCC1 cells. n = 4. ( g ) In vivo limiting dilution assay conducted to examine the effects of AFF4 depletion on tumor-initiation capacity of SCC1 cells in vivo . n = 6 for each group. Total mice, 48. ( h ) Quantification of formed tumors in g. ( i , j ) Flow cytometry assay performed to examine the effects of AFF4 overexpression on ALDH activity of SCC1 cells (i) and SCC23 cells (j). ( k ) Quantification of ALDH + cells in i and j. n = 3. ( l ) Sphere formation assay conducted to examine the effects of AFF4 overexpression on sphere formation capacity of SCC1 cells and SCC23 cells. Bar indicates 200 μm. ( m ) Quantification of formed spheres (diameter exceeding 10 µm) in k. n = 3. ( n ) qPCR analysis of the effects of AFF4 overexpression on expression of BMI1 , CD44 , NANOG and SOX2 in SCC1 cells. n = 4. * P -value
    Figure Legend Snippet: AFF4 enhances tumor-initiation capacity of HNSCC cells. ( a , b ) Flow cytometry assay performed to examine the effects of AFF4 depletion on ALDH activity of SCC1 cells (a) and SCC23 cells (b). Cells stained with ALDH+DEAB, an ALDH inhibitor, served as the negative control. ( c ) Quantification of ALDH + cells in a and b. n = 3. ( d ) Sphere formation assay conducted to examine the effects of AFF4 depletion on sphere formation capacity of SCC1 cells and SCC23 cells. Bar indicates 200 μm. ( e ) Quantification of formed spheres (diameter exceeding 10 µm) in d. n = 3. ( f ) qPCR analysis of the effects of AFF4 depletion on expression of BMI1 , CD44 , NANOG and SOX2 in SCC1 cells. n = 4. ( g ) In vivo limiting dilution assay conducted to examine the effects of AFF4 depletion on tumor-initiation capacity of SCC1 cells in vivo . n = 6 for each group. Total mice, 48. ( h ) Quantification of formed tumors in g. ( i , j ) Flow cytometry assay performed to examine the effects of AFF4 overexpression on ALDH activity of SCC1 cells (i) and SCC23 cells (j). ( k ) Quantification of ALDH + cells in i and j. n = 3. ( l ) Sphere formation assay conducted to examine the effects of AFF4 overexpression on sphere formation capacity of SCC1 cells and SCC23 cells. Bar indicates 200 μm. ( m ) Quantification of formed spheres (diameter exceeding 10 µm) in k. n = 3. ( n ) qPCR analysis of the effects of AFF4 overexpression on expression of BMI1 , CD44 , NANOG and SOX2 in SCC1 cells. n = 4. * P -value

    Techniques Used: Flow Cytometry, Cytometry, Activity Assay, Staining, Negative Control, Tube Formation Assay, Real-time Polymerase Chain Reaction, Expressing, In Vivo, Limiting Dilution Assay, Mouse Assay, Over Expression

    Overexpression of SOX2 rescued the inhibitory effects on HNSCC cell behavior induced by AFF4 depletion. ( a ) Western blot analysis of SOX2 expression in SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown at 48 h post transfection. n = 3. ( b , c ) MTT assay for cell proliferation in SOX2-overexpressing SCC1 cells (b) and SCC23 cells (c) in response to AFF4 knockdown at 3, 5 days. n = 3 at each time point. ( d ) Flow cytometry assay for ALDH activity of SOX2-overexpressing SCC1 cells in response to AFF4 knockdown at 48 h post transfection. n = 3. ( e ) Sphere formation assay conducted to evaluate sphere formation capacity of SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown after 2 weeks of culture in suspension. Bar indicates 200 μm. n = 3. ( f ) Would healing assay for migration activity of SOX2-overexpressing SCC1 cells in response to AFF4 knockdown. n = 3. ( g , h ) Transwell assay for invasion activity SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown. n = 3. * P -value
    Figure Legend Snippet: Overexpression of SOX2 rescued the inhibitory effects on HNSCC cell behavior induced by AFF4 depletion. ( a ) Western blot analysis of SOX2 expression in SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown at 48 h post transfection. n = 3. ( b , c ) MTT assay for cell proliferation in SOX2-overexpressing SCC1 cells (b) and SCC23 cells (c) in response to AFF4 knockdown at 3, 5 days. n = 3 at each time point. ( d ) Flow cytometry assay for ALDH activity of SOX2-overexpressing SCC1 cells in response to AFF4 knockdown at 48 h post transfection. n = 3. ( e ) Sphere formation assay conducted to evaluate sphere formation capacity of SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown after 2 weeks of culture in suspension. Bar indicates 200 μm. n = 3. ( f ) Would healing assay for migration activity of SOX2-overexpressing SCC1 cells in response to AFF4 knockdown. n = 3. ( g , h ) Transwell assay for invasion activity SOX2-overexpressing SCC1 and SCC23 cells in response to AFF4 knockdown. n = 3. * P -value

    Techniques Used: Over Expression, Western Blot, Expressing, Transfection, MTT Assay, Flow Cytometry, Cytometry, Activity Assay, Tube Formation Assay, Migration, Transwell Assay

    6) Product Images from "hESC Expansion and Stemness Are Independent of Connexin Forty-Three-Mediated Intercellular Communication between hESCs and hASC Feeder Cells"

    Article Title: hESC Expansion and Stemness Are Independent of Connexin Forty-Three-Mediated Intercellular Communication between hESCs and hASC Feeder Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0069175

    Culture of hESCs on Cx43 siRNA-treated hASCs. Compared with treatment of hASCs with control or scrambled-siRNA, the Cx43-siRNA treatment of hASC feeder cells did not alter adhesion (A), colony growth (n = 20; B), gene expression (i.e., OCT4 , SOX2 , and NANOG ) (C), AP-positive colony numbers (D), OCT4 expression (E), or chromosomal stability (F) of co-cultured hESCs. Nuclei were counterstained with DAPI (E). All data are shown as the mean ± the SD. (n = 4; # , p > 0.05). Scale bar, 100 μm.
    Figure Legend Snippet: Culture of hESCs on Cx43 siRNA-treated hASCs. Compared with treatment of hASCs with control or scrambled-siRNA, the Cx43-siRNA treatment of hASC feeder cells did not alter adhesion (A), colony growth (n = 20; B), gene expression (i.e., OCT4 , SOX2 , and NANOG ) (C), AP-positive colony numbers (D), OCT4 expression (E), or chromosomal stability (F) of co-cultured hESCs. Nuclei were counterstained with DAPI (E). All data are shown as the mean ± the SD. (n = 4; # , p > 0.05). Scale bar, 100 μm.

    Techniques Used: Expressing, Cell Culture

    Effect of Cx43 downregulation on hESC stemness. Downregulation of Cx43 and stemness gene ( OCT4 , SOX2 , and NANOG ) expression in Cx43-siRNA-treated hESCs was confirmed by qRT-PCR after 2 days culture (A) and Western blot analysis after 4 days culture (B). The percentage of AP-positive colonies formed by hESCs following Cx43-siRNA treatment was significantly lower than the percentage of AP-positive colonies formed by hESCs following control or scrambled-siRNA treatment (C). Immunocytochemical analysis demonstrated that OCT4 and Cx43 protein expression was reduced in Cx43-siRNA-treated hESCs versus control or scrambled-siRNA-treated hESCs (D). (C) and (D) were evaluated after 5 days culture. All data are shown as the mean ± the SD (n = 4; *, p
    Figure Legend Snippet: Effect of Cx43 downregulation on hESC stemness. Downregulation of Cx43 and stemness gene ( OCT4 , SOX2 , and NANOG ) expression in Cx43-siRNA-treated hESCs was confirmed by qRT-PCR after 2 days culture (A) and Western blot analysis after 4 days culture (B). The percentage of AP-positive colonies formed by hESCs following Cx43-siRNA treatment was significantly lower than the percentage of AP-positive colonies formed by hESCs following control or scrambled-siRNA treatment (C). Immunocytochemical analysis demonstrated that OCT4 and Cx43 protein expression was reduced in Cx43-siRNA-treated hESCs versus control or scrambled-siRNA-treated hESCs (D). (C) and (D) were evaluated after 5 days culture. All data are shown as the mean ± the SD (n = 4; *, p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    7) Product Images from "Establishment and Characterization of a Human Small Cell Osteosarcoma Cancer Stem Cell Line: A New Possible In Vitro Model for Discovering Small Cell Osteosarcoma Biology"

    Article Title: Establishment and Characterization of a Human Small Cell Osteosarcoma Cancer Stem Cell Line: A New Possible In Vitro Model for Discovering Small Cell Osteosarcoma Biology

    Journal: Stem Cells International

    doi: 10.1155/2016/3042198

    Immunofluorescence staining of ESCs markers. Immunofluorescence staining of Nanog (a), POU5F1 (c), SOX2 (e), KLF4 (g), and LIN-28A (i) of the cell line of OSA3-CSCs. (b, d, f, h, and j) Negative controls, respectively, for each marker. LSCM in conventional colors: green for ESCs and pink for nuclei. Original magnifications: 20x. Bar size: 100 µ m.
    Figure Legend Snippet: Immunofluorescence staining of ESCs markers. Immunofluorescence staining of Nanog (a), POU5F1 (c), SOX2 (e), KLF4 (g), and LIN-28A (i) of the cell line of OSA3-CSCs. (b, d, f, h, and j) Negative controls, respectively, for each marker. LSCM in conventional colors: green for ESCs and pink for nuclei. Original magnifications: 20x. Bar size: 100 µ m.

    Techniques Used: Immunofluorescence, Staining, Marker

    8) Product Images from "Progesterone Receptor Membrane Component 1 suppresses the p53 and Wnt/β-catenin pathways to promote human pluripotent stem cell self-renewal"

    Article Title: Progesterone Receptor Membrane Component 1 suppresses the p53 and Wnt/β-catenin pathways to promote human pluripotent stem cell self-renewal

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21322-z

    Expression of 108-B6 and 4A68 antigens is localized to undifferentiated and pluripotent hPSCs. ( a ) Multi-color flow cytometric analysis of hPSCs. H9 hPSCs were stained with MAbs (108-B6 or 4A68) and PE-conjugated anti-mouse IgG. The cells were then stained with anti-TRA-1-81 or anti-SSEA3 antibodies followed by incubation with FITC-conjugated anti-mouse IgM or anti-rat IgM, respectively, before analysis. ( b ) Multi-color intracellular flow cytometric analysis of hPSCs. H9 hPSCs were stained with MAbs 108-B6 or 4A68 and PE-conjugated anti-mouse IgG. The cells were fixed with 2% PFA and incubated in PBA containing 0.5% saponin. The cells were then stained with anti-OCT4 or anti-SOX2 antibodies followed by incubation with Alexa 488-cojugated anti-rabbit IgG before analysis.
    Figure Legend Snippet: Expression of 108-B6 and 4A68 antigens is localized to undifferentiated and pluripotent hPSCs. ( a ) Multi-color flow cytometric analysis of hPSCs. H9 hPSCs were stained with MAbs (108-B6 or 4A68) and PE-conjugated anti-mouse IgG. The cells were then stained with anti-TRA-1-81 or anti-SSEA3 antibodies followed by incubation with FITC-conjugated anti-mouse IgM or anti-rat IgM, respectively, before analysis. ( b ) Multi-color intracellular flow cytometric analysis of hPSCs. H9 hPSCs were stained with MAbs 108-B6 or 4A68 and PE-conjugated anti-mouse IgG. The cells were fixed with 2% PFA and incubated in PBA containing 0.5% saponin. The cells were then stained with anti-OCT4 or anti-SOX2 antibodies followed by incubation with Alexa 488-cojugated anti-rabbit IgG before analysis.

    Techniques Used: Expressing, Flow Cytometry, Staining, Incubation

    9) Product Images from "Expression of Pluripotent Stem Cell Reprogramming Factors by Prostate Tumor Initiating Cells"

    Article Title: Expression of Pluripotent Stem Cell Reprogramming Factors by Prostate Tumor Initiating Cells

    Journal: The Journal of urology

    doi: 10.1016/j.juro.2009.12.092

    Immunohistochemical staining for OCT3/4 and SOX2 in human prostate cancer tissues. A , representative results of at least 2 independent experiments reveal immunostaining for OCT3/4 and SOX2 using prostate tissue arrays. Neg ., negative. Int ., intermediate. Brown areas indicate positive nuclear staining. Reduced from ×20. Scale bar represents 70 µm. Insets, reduced from ×40. B and C , classification of different GS samples based on staining intensity. GS 5–6 and 7–8 tissues were significantly different than normal ( N ) and BPH tissue (2-tailed Mann-Whitney rank test p
    Figure Legend Snippet: Immunohistochemical staining for OCT3/4 and SOX2 in human prostate cancer tissues. A , representative results of at least 2 independent experiments reveal immunostaining for OCT3/4 and SOX2 using prostate tissue arrays. Neg ., negative. Int ., intermediate. Brown areas indicate positive nuclear staining. Reduced from ×20. Scale bar represents 70 µm. Insets, reduced from ×40. B and C , classification of different GS samples based on staining intensity. GS 5–6 and 7–8 tissues were significantly different than normal ( N ) and BPH tissue (2-tailed Mann-Whitney rank test p

    Techniques Used: Immunohistochemistry, Staining, Immunostaining, MANN-WHITNEY

    Representative results show identification of stem cell-like tumor cells with pluripotent stem cell reprogramming factors in 3 independent experiments in prostate cancer cell lines. A and B , RT-PCR or Western blot detected OCT3/4, SOX2, Nanog, c-Myc and Klf4 expression in DU145 and PC3 cell lines with human embryonic stem cells ( hESC ) as control. Data were normalized to β-actin. C , DU145 and PC3 cells were immunostained for OCT3/4 (red areas), SOX2 (green areas) and DAPI (blue areas). Arrow indicates merged OCT3/4 and SOX2 staining ( Merge ). Scale bar represents 20 µm. Reduced from ×40.
    Figure Legend Snippet: Representative results show identification of stem cell-like tumor cells with pluripotent stem cell reprogramming factors in 3 independent experiments in prostate cancer cell lines. A and B , RT-PCR or Western blot detected OCT3/4, SOX2, Nanog, c-Myc and Klf4 expression in DU145 and PC3 cell lines with human embryonic stem cells ( hESC ) as control. Data were normalized to β-actin. C , DU145 and PC3 cells were immunostained for OCT3/4 (red areas), SOX2 (green areas) and DAPI (blue areas). Arrow indicates merged OCT3/4 and SOX2 staining ( Merge ). Scale bar represents 20 µm. Reduced from ×40.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Staining

    Isolation of stem cell-like prostate tumor cells. A , to identify surface markers for isolating stem cell-like cells from prostate cell lines DU145 and PC3 cells were immunostained for OCT3/4 (red areas), E-cadherin ( Ecad ) (green areas) and DAPI (blue areas). OCT3/4 and E-cadherin staining was also merged (arrow). Reduced from ×40. Scale bar represents 20 µm. B and C , for phenotypic analysis of DU145 and PC3 cells using double staining with E-cadherin and CD44 or integrin-α2β1 cells were gated on E-cadherin + (green curves) or E-cadherin − (blue curves) population. FITC , fluorescein isothiocyanate. D and E , flow cytometry ( FCS ) analysis of DU145 and PC3 cells reveals E-cadherin ( E-cad ) expression. Isotype matched controls were used to set analysis gates for E-cadherin cell sorting. F , representative results of 3 independent experiments show that RT-PCR identified OCT3/4, SOX2, Nanog, c-Myc and Klf4 expression in E-cadherin + and E-cadherin − cells isolated from DU145 and PC3 cells. Data were normalized to β-actin.
    Figure Legend Snippet: Isolation of stem cell-like prostate tumor cells. A , to identify surface markers for isolating stem cell-like cells from prostate cell lines DU145 and PC3 cells were immunostained for OCT3/4 (red areas), E-cadherin ( Ecad ) (green areas) and DAPI (blue areas). OCT3/4 and E-cadherin staining was also merged (arrow). Reduced from ×40. Scale bar represents 20 µm. B and C , for phenotypic analysis of DU145 and PC3 cells using double staining with E-cadherin and CD44 or integrin-α2β1 cells were gated on E-cadherin + (green curves) or E-cadherin − (blue curves) population. FITC , fluorescein isothiocyanate. D and E , flow cytometry ( FCS ) analysis of DU145 and PC3 cells reveals E-cadherin ( E-cad ) expression. Isotype matched controls were used to set analysis gates for E-cadherin cell sorting. F , representative results of 3 independent experiments show that RT-PCR identified OCT3/4, SOX2, Nanog, c-Myc and Klf4 expression in E-cadherin + and E-cadherin − cells isolated from DU145 and PC3 cells. Data were normalized to β-actin.

    Techniques Used: Isolation, Staining, Double Staining, Flow Cytometry, Cytometry, Expressing, FACS, Reverse Transcription Polymerase Chain Reaction

    OCT3/4 or SOX2 knockdown tumorigenicity in DU145 and PC3 cells. A and B , Western blot shows decreased OCT3/4 or SOX2 protein levels in human DU145 and PC3 prostate cancer cells transfected with shRNA. Unsorted DU145 (1 × 10 5 ) or PC3 (3 × 10 5 ) cells were subcutaneously injected in SCID mice. C and D , mean ± SD tumor volume after OCT 3/4, SOX2 or control shRNA treatment.
    Figure Legend Snippet: OCT3/4 or SOX2 knockdown tumorigenicity in DU145 and PC3 cells. A and B , Western blot shows decreased OCT3/4 or SOX2 protein levels in human DU145 and PC3 prostate cancer cells transfected with shRNA. Unsorted DU145 (1 × 10 5 ) or PC3 (3 × 10 5 ) cells were subcutaneously injected in SCID mice. C and D , mean ± SD tumor volume after OCT 3/4, SOX2 or control shRNA treatment.

    Techniques Used: Western Blot, Transfection, shRNA, Injection, Mouse Assay

    10) Product Images from "Glycyrrhizic Acid-Induced Differentiation Repressed Stemness in Hepatocellular Carcinoma by Targeting c-Jun N-Terminal Kinase 1"

    Article Title: Glycyrrhizic Acid-Induced Differentiation Repressed Stemness in Hepatocellular Carcinoma by Targeting c-Jun N-Terminal Kinase 1

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2019.01431

    GA suppresses tumor growth and enhances the antitumor effect of sorafenib. Images of tumors (A) , tumor volumes (B) , and body weight (C) of PLC/PRF/5 transfected with shNC or shJNK1 in BALB/c nu/nu mice treated with 100 mg/kg GA or saline as control. (D) IHC staining indicates the expression of CSC markers (SOX2 and OCT4) and differentiation markers (AFP and HEPPAR1) in tumors. (E) IHC analysis of SOX2, OCT4, AFP, and HEPPAR1 in tumors (one-way ANOVA; *** P
    Figure Legend Snippet: GA suppresses tumor growth and enhances the antitumor effect of sorafenib. Images of tumors (A) , tumor volumes (B) , and body weight (C) of PLC/PRF/5 transfected with shNC or shJNK1 in BALB/c nu/nu mice treated with 100 mg/kg GA or saline as control. (D) IHC staining indicates the expression of CSC markers (SOX2 and OCT4) and differentiation markers (AFP and HEPPAR1) in tumors. (E) IHC analysis of SOX2, OCT4, AFP, and HEPPAR1 in tumors (one-way ANOVA; *** P

    Techniques Used: Planar Chromatography, Transfection, Mouse Assay, Immunohistochemistry, Staining, Expressing

    11) Product Images from "RhoGDIα suppresses self-renewal and tumorigenesis of glioma stem cells"

    Article Title: RhoGDIα suppresses self-renewal and tumorigenesis of glioma stem cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.11423

    The RhoGDIα expression in GSCs ( A ) FASC analysis of CD133 percentage before magnetic sorting. ( B ) FASC analysis of CD133 percentage after sorting of CD133 − and CD133 + cells. CD133 + cells were isolated from GSC2 and GSC5 by magnetic beads, the percentage of CD133 + cells was determined by FASC analysis relative to cells labeled with IgG isotype control antibody. ( C ) Immunoblot analysis of RhoGDIα protein level in matched CD133 − and CD133 + cell isolated from GSC2 and GSC5. ( D ) Immunoblot analysis of RhoGDIα, stem cell marker (SOX2) and differentiation markers (GFAP and Tuj1) during GSC2 differentiation. The numbers represent gray value relative to Actin. ( E ) Immunoblot analysis of RhoGDIα and stem cell markers (Nestin, SOX2 and Bmi1) in U87MG and U87MG-SLC cell. The numbers represent gray value relative to Actin.
    Figure Legend Snippet: The RhoGDIα expression in GSCs ( A ) FASC analysis of CD133 percentage before magnetic sorting. ( B ) FASC analysis of CD133 percentage after sorting of CD133 − and CD133 + cells. CD133 + cells were isolated from GSC2 and GSC5 by magnetic beads, the percentage of CD133 + cells was determined by FASC analysis relative to cells labeled with IgG isotype control antibody. ( C ) Immunoblot analysis of RhoGDIα protein level in matched CD133 − and CD133 + cell isolated from GSC2 and GSC5. ( D ) Immunoblot analysis of RhoGDIα, stem cell marker (SOX2) and differentiation markers (GFAP and Tuj1) during GSC2 differentiation. The numbers represent gray value relative to Actin. ( E ) Immunoblot analysis of RhoGDIα and stem cell markers (Nestin, SOX2 and Bmi1) in U87MG and U87MG-SLC cell. The numbers represent gray value relative to Actin.

    Techniques Used: Expressing, Isolation, Magnetic Beads, Labeling, Marker

    12) Product Images from "A post-transcriptional program coordinated by CSDE1 prevents intrinsic neural differentiation of human embryonic stem cells"

    Article Title: A post-transcriptional program coordinated by CSDE1 prevents intrinsic neural differentiation of human embryonic stem cells

    Journal: Nature Communications

    doi: 10.1038/s41467-017-01744-5

    CSDE1 prevents neural differentiation of hESCs. a Brightfield images of H9 hESCs. Knockdown of CSDE1 results in the proliferation of flattened and elongated cells that grew in monolayer colonies with reduced cell contact. Furthermore, loss of CSDE1 induces a spontaneous differentiation into neuronal cells. Scale bar represents 250 μm. b Percentage of alkaline phosphatase (AP)-positive colonies after five days of culturing without removing differentiated cells. Graph represents the mean ± s.e.m. of the percentage observed in four independent experiments (we assessed approximately 150 total colonies in each independent experiment). c qPCR analysis of hESCs cultured for five days without removal of differentiated cells. Graph (relative expression to non-targeting (NT) shRNA) represents the mean ± s.e.m. of four independent experiments with three biological replicates. d Western blot analysis with antibodies to CSDE1, OCT4 and SOX2. β-actin is the loading control. hESCs were grown for five days without removing differentiated cells. e Immunocytochemistry of H9 hESCs grown for five days without removal of differentiated cells. OCT4, PAX6, and Hoechst staining were used as markers of pluripotency, neuroectodermal differentiation, and nuclei, respectively. Scale bar represents 20 μm. f – j Graphs represent the percentage (mean ± s.e.m.) of OCT4 and PAX6-positive cells/total nuclei after five days in culture without removal of differentiated cells: f H9 hESCs, n = 3 independent experiments, 550–700 total cells per experiment; g H9 hESCs, n = 3, 420–1000 cells per experiment; h H1 hESCs, n = 3, 290–960 cells per experiment; i HUES9 hESCs, n = 3, 320–710 cells per experiment; j HUES6 hESCs, n = 3, 200–550 cells per experiment. KD = CSDE1 knockdown. KO = CSDE1 knockout. All the statistical comparisons were made by Student’s t-test for unpaired samples. P -value: *( P
    Figure Legend Snippet: CSDE1 prevents neural differentiation of hESCs. a Brightfield images of H9 hESCs. Knockdown of CSDE1 results in the proliferation of flattened and elongated cells that grew in monolayer colonies with reduced cell contact. Furthermore, loss of CSDE1 induces a spontaneous differentiation into neuronal cells. Scale bar represents 250 μm. b Percentage of alkaline phosphatase (AP)-positive colonies after five days of culturing without removing differentiated cells. Graph represents the mean ± s.e.m. of the percentage observed in four independent experiments (we assessed approximately 150 total colonies in each independent experiment). c qPCR analysis of hESCs cultured for five days without removal of differentiated cells. Graph (relative expression to non-targeting (NT) shRNA) represents the mean ± s.e.m. of four independent experiments with three biological replicates. d Western blot analysis with antibodies to CSDE1, OCT4 and SOX2. β-actin is the loading control. hESCs were grown for five days without removing differentiated cells. e Immunocytochemistry of H9 hESCs grown for five days without removal of differentiated cells. OCT4, PAX6, and Hoechst staining were used as markers of pluripotency, neuroectodermal differentiation, and nuclei, respectively. Scale bar represents 20 μm. f – j Graphs represent the percentage (mean ± s.e.m.) of OCT4 and PAX6-positive cells/total nuclei after five days in culture without removal of differentiated cells: f H9 hESCs, n = 3 independent experiments, 550–700 total cells per experiment; g H9 hESCs, n = 3, 420–1000 cells per experiment; h H1 hESCs, n = 3, 290–960 cells per experiment; i HUES9 hESCs, n = 3, 320–710 cells per experiment; j HUES6 hESCs, n = 3, 200–550 cells per experiment. KD = CSDE1 knockdown. KO = CSDE1 knockout. All the statistical comparisons were made by Student’s t-test for unpaired samples. P -value: *( P

    Techniques Used: Real-time Polymerase Chain Reaction, Cell Culture, Expressing, shRNA, Western Blot, Immunocytochemistry, Staining, Knock-Out

    13) Product Images from "Developmental history provides a roadmap for the emergence of tumor plasticity"

    Article Title: Developmental history provides a roadmap for the emergence of tumor plasticity

    Journal: Developmental cell

    doi: 10.1016/j.devcel.2018.02.024

    Concurrent loss of Nkx2-1 and gain of oncogenes results in conversion of lung epithelium into foregut-like or mid/hindgut-like tissues in organoid cultures independent of stromal cells A, Schematic representation of the isolation and organoid culture of lung epithelial cells from Sox2-creER/SIG f/f ::Nkx2-1 f/f and Sox2-creER/KrasG12D::Nkx2-1 f/f mice. B, Immunostaining for mid/hindgut markers KRT20 (green), HNF4A or SOX9 (red), and lung epithelial marker Nkx2-1 (gray) on organoid sections derived from Sox2-creER/KrasG12D:Nkx2-1 f/f . C, Immunostaining for foregut squamous epithelial markers KRT5 or TP63 (green), SOX2 (red), and lung epithelial marker Nkx2-1 (gray) on organoid sections derived from Sox2-creER/SIG f/f :Nkx2-1 f/f . D, Colony forming efficiency of Sox2-Kras and Sox2-Kras/Nkx2-1 f/f cells was not significantly different. E, Percentage of Ki67 + cells was significantly higher in Sox2-Kras/Nkx2-1 f/f cells than in Sox2-Kras cells (two-sided t test, * P
    Figure Legend Snippet: Concurrent loss of Nkx2-1 and gain of oncogenes results in conversion of lung epithelium into foregut-like or mid/hindgut-like tissues in organoid cultures independent of stromal cells A, Schematic representation of the isolation and organoid culture of lung epithelial cells from Sox2-creER/SIG f/f ::Nkx2-1 f/f and Sox2-creER/KrasG12D::Nkx2-1 f/f mice. B, Immunostaining for mid/hindgut markers KRT20 (green), HNF4A or SOX9 (red), and lung epithelial marker Nkx2-1 (gray) on organoid sections derived from Sox2-creER/KrasG12D:Nkx2-1 f/f . C, Immunostaining for foregut squamous epithelial markers KRT5 or TP63 (green), SOX2 (red), and lung epithelial marker Nkx2-1 (gray) on organoid sections derived from Sox2-creER/SIG f/f :Nkx2-1 f/f . D, Colony forming efficiency of Sox2-Kras and Sox2-Kras/Nkx2-1 f/f cells was not significantly different. E, Percentage of Ki67 + cells was significantly higher in Sox2-Kras/Nkx2-1 f/f cells than in Sox2-Kras cells (two-sided t test, * P

    Techniques Used: Isolation, Mouse Assay, Immunostaining, Marker, Derivative Assay

    Alveolar cells, but not airway cells, convert into alternate gut-like cell fates after loss of the lung lineage-specifying transcription factor Nkx2-1 A, Immunostaining on sections from Sox2-Nkx2-1 f/f mice (n = 3). Alcian Blue and Fast Red staining (left). Enlargements show the black boxed regions at higher magnification. Immunofluorescence for TFF2 (green), MUC5AC (red), and NKX2-1 (white) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). Nuclei, DAPI (blue). Black arrows, areas of mucous differentiation. B, Quantification of Nkx2-1 + cells normalized to total Sox2 + cells in the airways of Sox2-Nkx2-1 f/f mice (n = 3). Mice treated with tamoxifen had significantly fewer Nkx2-1 + cells (two-sided t test, ** P
    Figure Legend Snippet: Alveolar cells, but not airway cells, convert into alternate gut-like cell fates after loss of the lung lineage-specifying transcription factor Nkx2-1 A, Immunostaining on sections from Sox2-Nkx2-1 f/f mice (n = 3). Alcian Blue and Fast Red staining (left). Enlargements show the black boxed regions at higher magnification. Immunofluorescence for TFF2 (green), MUC5AC (red), and NKX2-1 (white) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). Nuclei, DAPI (blue). Black arrows, areas of mucous differentiation. B, Quantification of Nkx2-1 + cells normalized to total Sox2 + cells in the airways of Sox2-Nkx2-1 f/f mice (n = 3). Mice treated with tamoxifen had significantly fewer Nkx2-1 + cells (two-sided t test, ** P

    Techniques Used: Immunostaining, Mouse Assay, Staining, Immunofluorescence

    Concurrent loss of Nkx2-1 and oncogene activation leads to acquisition of mid/hindgut-like cell fates in lung mucinous adenocarcinomas A, Immunostaining on sections from Sox2-Kras mice (n = 3). Alcian Blue and Fast Red staining (left). Immunofluorescence for TFF2 (green) and SCGB1A1 (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). B, Immunostaining on sections from Sox2-Kras-Nkx2-1 f/f mice (n = 4). Alcian Blue and Fast Red staining (left). Immunofluorescence for TFF2 (green) and MUC5AC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). C, Immunostaining on sections from Sftpc-Kras mice (n = 6). Alcian Blue and Fast Red staining (left). Immunostaining for TFF2 (green) and SFTPC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). D, Immunostaining on sections from Sftpc-Kras-Nkx2-1 f/f mice (n = 4). Alcian Blue and Fast Red staining (left). Immunostaining for TFF2 (green) and MUC5AC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). Nuclei, DAPI (blue). Scale bars, 20 μm. E, Immunostaining on sections from Sox2-Kras-Nkx2-1 +/+ /LSL-YFP mice for YFP/GFP (green), CD31 (red), and FSP1 (purple). Nuclei, DAPI (blue). F, Quantification of Ki67 + proliferating cells among total DAPI + cells in Sftpc-Kras (blue) (n = 3) and Sftpc-Kras-Nkx2-1 f/f (green) (n = 3) mice. The proportion of Ki67 + cells was not significantly different between the two genotypes (two-sided t test, P .
    Figure Legend Snippet: Concurrent loss of Nkx2-1 and oncogene activation leads to acquisition of mid/hindgut-like cell fates in lung mucinous adenocarcinomas A, Immunostaining on sections from Sox2-Kras mice (n = 3). Alcian Blue and Fast Red staining (left). Immunofluorescence for TFF2 (green) and SCGB1A1 (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). B, Immunostaining on sections from Sox2-Kras-Nkx2-1 f/f mice (n = 4). Alcian Blue and Fast Red staining (left). Immunofluorescence for TFF2 (green) and MUC5AC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). C, Immunostaining on sections from Sftpc-Kras mice (n = 6). Alcian Blue and Fast Red staining (left). Immunostaining for TFF2 (green) and SFTPC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). D, Immunostaining on sections from Sftpc-Kras-Nkx2-1 f/f mice (n = 4). Alcian Blue and Fast Red staining (left). Immunostaining for TFF2 (green) and MUC5AC (red) (middle), HNF4α (green), PDX1 (red), and NKX2-1 (white) (right). Nuclei, DAPI (blue). Scale bars, 20 μm. E, Immunostaining on sections from Sox2-Kras-Nkx2-1 +/+ /LSL-YFP mice for YFP/GFP (green), CD31 (red), and FSP1 (purple). Nuclei, DAPI (blue). F, Quantification of Ki67 + proliferating cells among total DAPI + cells in Sftpc-Kras (blue) (n = 3) and Sftpc-Kras-Nkx2-1 f/f (green) (n = 3) mice. The proportion of Ki67 + cells was not significantly different between the two genotypes (two-sided t test, P .

    Techniques Used: Activation Assay, Immunostaining, Mouse Assay, Staining, Immunofluorescence

    Loss of Nkx2-1 and gain of Sox2 induces conversion of lung epithelium into esophageal-like squamous epithelium in lung squamous cell carcinoma A, Staining on sections from Sox2-SIG f/+ mice (n = 4). Hematoxylin and eosin staining (left). Immunofluorescence for GFP (green), FOXJ1 (red) and CK6 (white) (middle), GFP (green), SCGB1A1 (red), and AcTub (white) (right). B, Quantification of GFP+ cells in Sox2-SIG f/+ (n = 3) lungs. Ciliated cells (blue), secretory cells (gray), squamous cells (orange), and all other cells (navy). C, Staining on sections from Sox2-Nkx2-1 f/f -SIG f/f mice (n = 8 mice). Hematoxylin and eosin staining (left). Immunostaining for GFP (green) and CK6 (red) (middle), SOX2 (green), CK6 (red), and NKX2-1 (white) (right). D, Quantification of GFP+ cells in Sox2-Nkx2-1 f/f -SIG f/f (n = 7; right) lungs. Ciliated cells (blue), secretory cells (gray), squamous cells (orange), and all other cells (navy). E, Immunostaining on sections from Sox2-SIG f/f -Nkx2-1 f/f mice for YFP/GFP (green), CD31 (red), and FSP1 (purple). Nuclei, DAPI (blue). F, Tumors derived from subcutaneous grafts were stained for SOX2 (red), GFP (green), and KRT14 (purple). G, Quantification of Ki67 + proliferating cells among total DAPI + cells. Sox2-Nkx2-1 f/f -SIG f/f mice had significantly more Ki67 + cells than Sox2-Nkx2-1 f/f mice (two-sided t test, * P
    Figure Legend Snippet: Loss of Nkx2-1 and gain of Sox2 induces conversion of lung epithelium into esophageal-like squamous epithelium in lung squamous cell carcinoma A, Staining on sections from Sox2-SIG f/+ mice (n = 4). Hematoxylin and eosin staining (left). Immunofluorescence for GFP (green), FOXJ1 (red) and CK6 (white) (middle), GFP (green), SCGB1A1 (red), and AcTub (white) (right). B, Quantification of GFP+ cells in Sox2-SIG f/+ (n = 3) lungs. Ciliated cells (blue), secretory cells (gray), squamous cells (orange), and all other cells (navy). C, Staining on sections from Sox2-Nkx2-1 f/f -SIG f/f mice (n = 8 mice). Hematoxylin and eosin staining (left). Immunostaining for GFP (green) and CK6 (red) (middle), SOX2 (green), CK6 (red), and NKX2-1 (white) (right). D, Quantification of GFP+ cells in Sox2-Nkx2-1 f/f -SIG f/f (n = 7; right) lungs. Ciliated cells (blue), secretory cells (gray), squamous cells (orange), and all other cells (navy). E, Immunostaining on sections from Sox2-SIG f/f -Nkx2-1 f/f mice for YFP/GFP (green), CD31 (red), and FSP1 (purple). Nuclei, DAPI (blue). F, Tumors derived from subcutaneous grafts were stained for SOX2 (red), GFP (green), and KRT14 (purple). G, Quantification of Ki67 + proliferating cells among total DAPI + cells. Sox2-Nkx2-1 f/f -SIG f/f mice had significantly more Ki67 + cells than Sox2-Nkx2-1 f/f mice (two-sided t test, * P

    Techniques Used: Staining, Mouse Assay, Immunofluorescence, Immunostaining, Derivative Assay

    Molecular and histologic features of human NSCLCs are reminiscent of embryonic gut endodermal patterning events A, Classification of non-small cell lung tumors (n = 1019) into three groups by expression of SOX2 and NKX2-1 . ADC (adenocarcinoma, n = 474), SCC (squamous cell carcinoma, n = 485), and MAD (mucinous adenocarcinoma, n = 60). B, Heat map of select differentially expressed genes. C, Immunohistochemistry for an SCC marker p40 (left) and an ADC marker NKX2-1 (right) on an adenosquamous lung tumor. Yellow dotted line indicates the boundary between SCC and ADC within a single tumor. D, Immunostaining on sections from human SCC for SOX2 (green), PAX9 (red), and NKX2-1(white) (left), CK6 (green) and CK1 (red) (middle), p63 (green) and CK14 (red) (right). E, Immunostaining on serial sections from human MADs. H E (top row, left), Alcian blue (top row, center), and TFF2 (green) (top row, right); HNF4α (green), PDX1 (red), and NKX2-1 (white) (middle row, left); CDX2 (green), SOX9 (red), and NKX2-1 (white) (middle row, center); PRSS1 (red) (middle row, right). CDX2 (green) and PDX1 (red) (bottom row, left); CDX2 (green), PDX1 (red), and MUC6 (cyan) (bottom row, center). Dotted lines correspond to boundaries in organ-specific differentiation in contiguous gut epithelia. Color-coded boxes in the top, middle and bottom panels correspond to specified regions on the H E image. F, .
    Figure Legend Snippet: Molecular and histologic features of human NSCLCs are reminiscent of embryonic gut endodermal patterning events A, Classification of non-small cell lung tumors (n = 1019) into three groups by expression of SOX2 and NKX2-1 . ADC (adenocarcinoma, n = 474), SCC (squamous cell carcinoma, n = 485), and MAD (mucinous adenocarcinoma, n = 60). B, Heat map of select differentially expressed genes. C, Immunohistochemistry for an SCC marker p40 (left) and an ADC marker NKX2-1 (right) on an adenosquamous lung tumor. Yellow dotted line indicates the boundary between SCC and ADC within a single tumor. D, Immunostaining on sections from human SCC for SOX2 (green), PAX9 (red), and NKX2-1(white) (left), CK6 (green) and CK1 (red) (middle), p63 (green) and CK14 (red) (right). E, Immunostaining on serial sections from human MADs. H E (top row, left), Alcian blue (top row, center), and TFF2 (green) (top row, right); HNF4α (green), PDX1 (red), and NKX2-1 (white) (middle row, left); CDX2 (green), SOX9 (red), and NKX2-1 (white) (middle row, center); PRSS1 (red) (middle row, right). CDX2 (green) and PDX1 (red) (bottom row, left); CDX2 (green), PDX1 (red), and MUC6 (cyan) (bottom row, center). Dotted lines correspond to boundaries in organ-specific differentiation in contiguous gut epithelia. Color-coded boxes in the top, middle and bottom panels correspond to specified regions on the H E image. F, .

    Techniques Used: Expressing, Immunohistochemistry, Marker, Immunostaining

    14) Product Images from "Human Cytomegalovirus Gene Expression in Long-Term Infected Glioma Stem Cells"

    Article Title: Human Cytomegalovirus Gene Expression in Long-Term Infected Glioma Stem Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0116178

    HCMV Infection Efficiency. A. U87 cells cultured on cover glass and treated with mock or TR (MOI = 2) were fixed with ethanol (72 hrs p.i.) and processed for IE1 immunofluorescence in conjunction with an Alexa 488-labeled secondary antibody. Nuclei were counterstained with DAPI. Bar = 50 um. B. GSC 387 grown as neurospheres were infected with TR (MOI = 2) and stained for IE1 and Sox2. Photomicrographs demonstrate co-localization of the two markers. Bar = 100 um. C. To quantitatively assess HCMV infection efficiency in GSC, 387 cells were cultured on laminin, treated with mock or TR (MOI = 2) and 72 h later processed for IE1 and Nestin immunofluorescence. Nestin is a marker of un-differentiation abundant in GSC. Nuclei were counterstained with DAPI. Rightmost panels show superimposed images of Nestin+, IE1+ cells. Bar = 100 um. D. Cell lines and GSC cultures treated as described above were counted using a fluorescence microscope fitted with a camera. Four low magnification fields were counted for each condition (repeated in duplicate coverslips). Percentage of positive cells was calculated by dividing the number of IE1+ cells to the total number of nuclei and multiplying by 100. Student t-test was used to compare the number of 387 IE1 positive cells at 5 weeks p.i. (green bars) with the number of T98G and U87 IE1 positive cells, respectively. For both viral strains, differences between IE1+ cells in GSC and cell lines were significant (p values displayed).
    Figure Legend Snippet: HCMV Infection Efficiency. A. U87 cells cultured on cover glass and treated with mock or TR (MOI = 2) were fixed with ethanol (72 hrs p.i.) and processed for IE1 immunofluorescence in conjunction with an Alexa 488-labeled secondary antibody. Nuclei were counterstained with DAPI. Bar = 50 um. B. GSC 387 grown as neurospheres were infected with TR (MOI = 2) and stained for IE1 and Sox2. Photomicrographs demonstrate co-localization of the two markers. Bar = 100 um. C. To quantitatively assess HCMV infection efficiency in GSC, 387 cells were cultured on laminin, treated with mock or TR (MOI = 2) and 72 h later processed for IE1 and Nestin immunofluorescence. Nestin is a marker of un-differentiation abundant in GSC. Nuclei were counterstained with DAPI. Rightmost panels show superimposed images of Nestin+, IE1+ cells. Bar = 100 um. D. Cell lines and GSC cultures treated as described above were counted using a fluorescence microscope fitted with a camera. Four low magnification fields were counted for each condition (repeated in duplicate coverslips). Percentage of positive cells was calculated by dividing the number of IE1+ cells to the total number of nuclei and multiplying by 100. Student t-test was used to compare the number of 387 IE1 positive cells at 5 weeks p.i. (green bars) with the number of T98G and U87 IE1 positive cells, respectively. For both viral strains, differences between IE1+ cells in GSC and cell lines were significant (p values displayed).

    Techniques Used: Infection, Cell Culture, Immunofluorescence, Labeling, Staining, Marker, Fluorescence, Microscopy

    15) Product Images from "Nanog interaction with the androgen receptor signaling axis induce ovarian cancer stem cell regulation: studies based on the CRISPR/Cas9 system"

    Article Title: Nanog interaction with the androgen receptor signaling axis induce ovarian cancer stem cell regulation: studies based on the CRISPR/Cas9 system

    Journal: Journal of Ovarian Research

    doi: 10.1186/s13048-018-0403-2

    Protein expression of other stem cell genes exhibits the same tendency as Nanog. a and b ) Oct4 and Sox2 expression increased in the GFP (+) cells compared with the GFP (−) cells according to western blot analysis. c and d ) Oct4 and Sox2 expression in the GFP (+) cells clearly increased under DHT treatment compared with ASC-J9 treatment based on western blot analysis, which was also consistent with the Nanog expression trend
    Figure Legend Snippet: Protein expression of other stem cell genes exhibits the same tendency as Nanog. a and b ) Oct4 and Sox2 expression increased in the GFP (+) cells compared with the GFP (−) cells according to western blot analysis. c and d ) Oct4 and Sox2 expression in the GFP (+) cells clearly increased under DHT treatment compared with ASC-J9 treatment based on western blot analysis, which was also consistent with the Nanog expression trend

    Techniques Used: Expressing, Western Blot

    16) Product Images from "Targeting BMI1+ Cancer Stem Cells Overcomes Chemoresistance and Inhibits Metastases in Squamous Cell Carcinoma"

    Article Title: Targeting BMI1+ Cancer Stem Cells Overcomes Chemoresistance and Inhibits Metastases in Squamous Cell Carcinoma

    Journal: Cell stem cell

    doi: 10.1016/j.stem.2017.02.003

    Bmi1 is expressed in a subset of cancer cells in HNSCC in vivo (A) Experimental design for tracing Bmi1 + cells in mouse HNSCC induced by 4NQO. (B) Representative image of tongue lesions 26 weeks after 4NQO treatment. Representative of at least three independent experiments. (C) Representative H E staining of adjacent normal tongue tissue (Adj normal), hyperplasia, papilloma and SCC. Tamoxifen was injected into 4NQO-treated Bmi CreER ;Rosa26 tdTomato mice. 24 hours after tamoxifen treatment, tongues were harvested and analyzed by immunostaining. The majority of Bmi1 + cells (red) resided within basal layer of SCC were stained by cytokeratin 5 (K5; green). Integrin α6 (α6; green) denotes the boundary of tumor epithelia and stroma. Scale bars, 50 μm. (D) Expression pattern of Bmi1 mRNA in both papilloma and SCC was detected by in situ hybridization. Scale bars, 50 μm. (E) Representative images of Bmi1 + cell-driven lineage tracing in primary HNSCC in vivo. Nuclei were stained with DAPI (blue). White dashed lines demarks tumor-stromal junction. (F) Immunostaining of Keratin14 (K14; green, left) or Keratin10 (K10; green, right) in mouse HNSCC. Scale bars, 50 μm. (G) Immunostaining of CD44 (green, left) or Sox2 (green, right) in mouse HNSCC. Scale bars, 50 μm. (H) A representative image of primary tumor cells labeled with EdU (green). Scale bars, 50 μm. (I) Quantification of EdU incorporation in Bmi1 + and Bmi1 − tumor cells. Values are mean ± SD. from one of three independent experiments at least. **p
    Figure Legend Snippet: Bmi1 is expressed in a subset of cancer cells in HNSCC in vivo (A) Experimental design for tracing Bmi1 + cells in mouse HNSCC induced by 4NQO. (B) Representative image of tongue lesions 26 weeks after 4NQO treatment. Representative of at least three independent experiments. (C) Representative H E staining of adjacent normal tongue tissue (Adj normal), hyperplasia, papilloma and SCC. Tamoxifen was injected into 4NQO-treated Bmi CreER ;Rosa26 tdTomato mice. 24 hours after tamoxifen treatment, tongues were harvested and analyzed by immunostaining. The majority of Bmi1 + cells (red) resided within basal layer of SCC were stained by cytokeratin 5 (K5; green). Integrin α6 (α6; green) denotes the boundary of tumor epithelia and stroma. Scale bars, 50 μm. (D) Expression pattern of Bmi1 mRNA in both papilloma and SCC was detected by in situ hybridization. Scale bars, 50 μm. (E) Representative images of Bmi1 + cell-driven lineage tracing in primary HNSCC in vivo. Nuclei were stained with DAPI (blue). White dashed lines demarks tumor-stromal junction. (F) Immunostaining of Keratin14 (K14; green, left) or Keratin10 (K10; green, right) in mouse HNSCC. Scale bars, 50 μm. (G) Immunostaining of CD44 (green, left) or Sox2 (green, right) in mouse HNSCC. Scale bars, 50 μm. (H) A representative image of primary tumor cells labeled with EdU (green). Scale bars, 50 μm. (I) Quantification of EdU incorporation in Bmi1 + and Bmi1 − tumor cells. Values are mean ± SD. from one of three independent experiments at least. **p

    Techniques Used: In Vivo, Staining, Injection, Mouse Assay, Immunostaining, Expressing, In Situ Hybridization, Labeling

    17) Product Images from "A High-Content Small Molecule Screen Identifies Sensitivity of Glioblastoma Stem Cells to Inhibition of Polo-Like Kinase 1"

    Article Title: A High-Content Small Molecule Screen Identifies Sensitivity of Glioblastoma Stem Cells to Inhibition of Polo-Like Kinase 1

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0077053

    BI 2536, a potent and selective PLK1 inhibitor, disrupts GNS cell proliferation but does not trigger astrocyte differentiation. (A)Dose responses to J101 and BI2536 for three different NS and GNS cell lines, assayed using pHH3 immunocytochemistry 24 h after treatment. (B) Cell cycle profiles of BI 2536-treated NS and GNS cells by flow cytometry confirm a clear increase in the proportion of cells in G2/M for GNS cells, but not normal NS cells. (C) Following BI 2536 treatment, GNS cells display reduced levels of SOX2 but do not have significantly altered levels of the neural stem cell marker SOX9 or upregulation of the astrocyte marker GFAP. (D) Mitotic shake-off was used to enrich for GNS cells in mitosis (confirmed by enrichment for Cyclin B1; left panel). Levels of SOX2 protein were compared in the population as a whole and those undergoing mitosis by immunoblotting. These results suggest a reduction of SOX2 occurs normally during mitosis.
    Figure Legend Snippet: BI 2536, a potent and selective PLK1 inhibitor, disrupts GNS cell proliferation but does not trigger astrocyte differentiation. (A)Dose responses to J101 and BI2536 for three different NS and GNS cell lines, assayed using pHH3 immunocytochemistry 24 h after treatment. (B) Cell cycle profiles of BI 2536-treated NS and GNS cells by flow cytometry confirm a clear increase in the proportion of cells in G2/M for GNS cells, but not normal NS cells. (C) Following BI 2536 treatment, GNS cells display reduced levels of SOX2 but do not have significantly altered levels of the neural stem cell marker SOX9 or upregulation of the astrocyte marker GFAP. (D) Mitotic shake-off was used to enrich for GNS cells in mitosis (confirmed by enrichment for Cyclin B1; left panel). Levels of SOX2 protein were compared in the population as a whole and those undergoing mitosis by immunoblotting. These results suggest a reduction of SOX2 occurs normally during mitosis.

    Techniques Used: Immunocytochemistry, Flow Cytometry, Cytometry, Marker

    18) Product Images from "Pranlukast Antagonizes CD49f and Reduces Stemness in Triple-Negative Breast Cancer Cells"

    Article Title: Pranlukast Antagonizes CD49f and Reduces Stemness in Triple-Negative Breast Cancer Cells

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S247730

    Effect of pranlukast on breast cancer cell stemness. ( A ) Experimental strategy. ( B ) Viable MDA-MB-231 cells previously exposed to pranlukast for 24 h were seeded to test their capacity to form mammospheres in the absence of the drug. Graphs show the mean number of mammospheres ± SD from three independent experiments. ( C ) Analysis of CD44 expression by flow cytometry. The annotated percentages correspond to CD44 hi population. A representative experiment from 2 is shown. ( D ) Representative Western blot analyzing SOX2 expression. Graph represents the mean ± SEM from three independent experiments. ( E ) Analysis of SOX2 promoter transactivation by luciferase assay. Values are mean ± SEM from four independent experiments. Scheme above graph shows the promoter size in the construct. Statistical significance in ( B ), ( D ), and ( E ) was determined by Dunnett´s test; P value
    Figure Legend Snippet: Effect of pranlukast on breast cancer cell stemness. ( A ) Experimental strategy. ( B ) Viable MDA-MB-231 cells previously exposed to pranlukast for 24 h were seeded to test their capacity to form mammospheres in the absence of the drug. Graphs show the mean number of mammospheres ± SD from three independent experiments. ( C ) Analysis of CD44 expression by flow cytometry. The annotated percentages correspond to CD44 hi population. A representative experiment from 2 is shown. ( D ) Representative Western blot analyzing SOX2 expression. Graph represents the mean ± SEM from three independent experiments. ( E ) Analysis of SOX2 promoter transactivation by luciferase assay. Values are mean ± SEM from four independent experiments. Scheme above graph shows the promoter size in the construct. Statistical significance in ( B ), ( D ), and ( E ) was determined by Dunnett´s test; P value

    Techniques Used: Multiple Displacement Amplification, Expressing, Flow Cytometry, Western Blot, Luciferase, Construct

    19) Product Images from "REST overexpression in mice causes deficits in spontaneous locomotion"

    Article Title: REST overexpression in mice causes deficits in spontaneous locomotion

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-29441-3

    Characterization of N-hREST mice. ( a ) Nestin-Cre x LSL-hREST ( N-hREST ) heterozygous pups were born dead and had smaller brains than the LSL-hREST control pups. Body, brain weight, and brain/body weight of E18.5 embryos are shown. Experiments were performed at least in triplicate. P values are shown in the figure. ( b , c ) REST expression in N-hREST mouse brains correlates with stemness in embryonic neural stem cells. Immunofluorescence analysis of E18.5 N-hREST and LSL-hREST control littermate mouse brains with antibodies against REST (using an antibody that preferentially recognizes hREST over mouse REST) and Nestin ( b ) and REST and SOX2 ( c ). Figures represent at least three different images. Scale bar for B and C = 100 μ.
    Figure Legend Snippet: Characterization of N-hREST mice. ( a ) Nestin-Cre x LSL-hREST ( N-hREST ) heterozygous pups were born dead and had smaller brains than the LSL-hREST control pups. Body, brain weight, and brain/body weight of E18.5 embryos are shown. Experiments were performed at least in triplicate. P values are shown in the figure. ( b , c ) REST expression in N-hREST mouse brains correlates with stemness in embryonic neural stem cells. Immunofluorescence analysis of E18.5 N-hREST and LSL-hREST control littermate mouse brains with antibodies against REST (using an antibody that preferentially recognizes hREST over mouse REST) and Nestin ( b ) and REST and SOX2 ( c ). Figures represent at least three different images. Scale bar for B and C = 100 μ.

    Techniques Used: Mouse Assay, Expressing, Immunofluorescence

    20) Product Images from "Different Effects of Human Umbilical Cord Mesenchymal Stem Cells on Glioblastoma Stem Cells by Direct Cell Interaction or Via Released Soluble Factors"

    Article Title: Different Effects of Human Umbilical Cord Mesenchymal Stem Cells on Glioblastoma Stem Cells by Direct Cell Interaction or Via Released Soluble Factors

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2017.00312

    Characterization of glioblastoma (GBM) cancer stem cells (CSCs). (A) Morphological appearance of CSCs grown as floating spheres in complete stem medium (Stem) and as monolayers after induction of differentiation in serum-containing medium (Diff.). (B) Representative immunofluorescence images of Sox2 (red nuclear staining) and GFAP (red cytoskeleton staining) expression in CSCs before and after induction of differentiation in serum-containing medium. Nuclei were counterstained with DAPI (blue). Scale, 100 μm. (C) Quantitative RT-PCR of GFAP mRNA levels in CSC1, CSC2 and CSC3 cells cultured in complete stem medium or after differentiation. Normalized values are referred to stem condition. Values are expressed as the mean of three experiments ± SD.
    Figure Legend Snippet: Characterization of glioblastoma (GBM) cancer stem cells (CSCs). (A) Morphological appearance of CSCs grown as floating spheres in complete stem medium (Stem) and as monolayers after induction of differentiation in serum-containing medium (Diff.). (B) Representative immunofluorescence images of Sox2 (red nuclear staining) and GFAP (red cytoskeleton staining) expression in CSCs before and after induction of differentiation in serum-containing medium. Nuclei were counterstained with DAPI (blue). Scale, 100 μm. (C) Quantitative RT-PCR of GFAP mRNA levels in CSC1, CSC2 and CSC3 cells cultured in complete stem medium or after differentiation. Normalized values are referred to stem condition. Values are expressed as the mean of three experiments ± SD.

    Techniques Used: Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Cell Culture

    21) Product Images from "Effect of histone deacetylase inhibitors trichostatin A and valproic acid on hair cell regeneration in zebrafish lateral line neuromasts"

    Article Title: Effect of histone deacetylase inhibitors trichostatin A and valproic acid on hair cell regeneration in zebrafish lateral line neuromasts

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2014.00382

    HDAC inhibitors significantly suppresses cell proliferation. (A and B) Lateral line SCs are stained with Sox2, and the BrdU antibody shows dividing cells in the neuromasts of zebrafish. (C and D) Quantification of the ratio of BrdU + SCs in control and TSA- or VPA-treated larvae at 24 h and 48 h after neomycin incubation. Bars are mean ± s.e.m. and n = total number of embryos. ** p
    Figure Legend Snippet: HDAC inhibitors significantly suppresses cell proliferation. (A and B) Lateral line SCs are stained with Sox2, and the BrdU antibody shows dividing cells in the neuromasts of zebrafish. (C and D) Quantification of the ratio of BrdU + SCs in control and TSA- or VPA-treated larvae at 24 h and 48 h after neomycin incubation. Bars are mean ± s.e.m. and n = total number of embryos. ** p

    Techniques Used: Staining, Incubation

    HDAC inhibitors significantly suppress SC production. (A and B) We treated larvae at 5 dpf with neomycin and monitored SC numbers over the following 2 days. Lateral line SCs are stained with Sox2 antibody. (C and D) Quantification of the number of SCs in control and TSA- or VPA-treated larvae at 24 h and 48 h after neomycin incubation. Bars are mean ± s.e.m. and n = total number of embryos. ** p
    Figure Legend Snippet: HDAC inhibitors significantly suppress SC production. (A and B) We treated larvae at 5 dpf with neomycin and monitored SC numbers over the following 2 days. Lateral line SCs are stained with Sox2 antibody. (C and D) Quantification of the number of SCs in control and TSA- or VPA-treated larvae at 24 h and 48 h after neomycin incubation. Bars are mean ± s.e.m. and n = total number of embryos. ** p

    Techniques Used: Staining, Incubation

    22) Product Images from "Remnant living cells that escape cell loss in late-stage tumors exhibit cancer stem cell-like characteristics"

    Article Title: Remnant living cells that escape cell loss in late-stage tumors exhibit cancer stem cell-like characteristics

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2012.136

    Expression of CSC-related markers in ex vivo isolated remnant living cells. ( a ) Oct4 mRNA and ( b ) Sox2 mRNA expression was compared between cultured and ex vivo isolated remnant living 4T1-PB-2R/PBase cells using qPCR. ( c ) Luciferase reporter-gene assay for investigating the Oct4 transcriptional activity in parental cells and isolated remnant living cells. The data represent the means of four independent experiments±S.D. * P
    Figure Legend Snippet: Expression of CSC-related markers in ex vivo isolated remnant living cells. ( a ) Oct4 mRNA and ( b ) Sox2 mRNA expression was compared between cultured and ex vivo isolated remnant living 4T1-PB-2R/PBase cells using qPCR. ( c ) Luciferase reporter-gene assay for investigating the Oct4 transcriptional activity in parental cells and isolated remnant living cells. The data represent the means of four independent experiments±S.D. * P

    Techniques Used: Expressing, Ex Vivo, Isolation, Cell Culture, Real-time Polymerase Chain Reaction, Luciferase, Reporter Gene Assay, Activity Assay

    23) Product Images from "Intermittent high oxygen influences the formation of neural retinal tissue from human embryonic stem cells"

    Article Title: Intermittent high oxygen influences the formation of neural retinal tissue from human embryonic stem cells

    Journal: Scientific Reports

    doi: 10.1038/srep29944

    High oxygen concentration facilitates neural retinal development similar to the status in vivo . ( a – f ) SOX2 and PAX6 double-staining of NE in ( a ) 38D 20% O 2 ; ( b ) 50D 20% O 2 ; ( c ) 62D 20% O 2 ; ( d ) 32D 40% O 2 ; ( e ) 52D 20% O 2 ; ( f ) 62D 40% O 2 . White arrows point at the PAX6-immunoreactive cells within the NE. ( g ) Ratio of PAX6-immunoreactive cells within the NE in 6 groups. The number of PAX6 positive cells within the NE in the high oxygen groups was much higher compared to the normal oxygen concentration groups. NE: Neuroectodermal epithelium.
    Figure Legend Snippet: High oxygen concentration facilitates neural retinal development similar to the status in vivo . ( a – f ) SOX2 and PAX6 double-staining of NE in ( a ) 38D 20% O 2 ; ( b ) 50D 20% O 2 ; ( c ) 62D 20% O 2 ; ( d ) 32D 40% O 2 ; ( e ) 52D 20% O 2 ; ( f ) 62D 40% O 2 . White arrows point at the PAX6-immunoreactive cells within the NE. ( g ) Ratio of PAX6-immunoreactive cells within the NE in 6 groups. The number of PAX6 positive cells within the NE in the high oxygen groups was much higher compared to the normal oxygen concentration groups. NE: Neuroectodermal epithelium.

    Techniques Used: Concentration Assay, In Vivo, Double Staining

    Cell proliferation within the NE requires oxygen support. ( a , c , e ) Tuj1 + Ki67, Tuj1 + NESTIN, Tuj1 + SOX2 double-stained NE in 62D 20% O 2 group. ( b , d , f ) Tuj1 + Ki67, Tuj1 + NESTIN, Tuj1 + SOX2 double-stained in 50D 40% O 2 group. White arrows in ( a , b ) point at the Ki67-positive proliferating cells in the NE of EB. The area between the two dotted lines in ( c , d ) represents the NESTIN-positive neural stem cells. The area between the two dotted lines in ( f ) represents the plate-ranged Tuj1-positive neurons. ( g , h ) Enlarged images of rosettes fusing at the edge of the EB in ( e ). ( i ) Enlarged Tuj1 (green) and SOX2 (red) double-stained images of typical rosettes fusing at the edge of EB in 50D 20% O 2 . White arrows in g , ( h , i ) point at the extroverting inner side of the rosettes. ( j – l ) ZO-1 staining of apical surface in NE. ( j ) An out-apical-side NE without forming rosettes. ( k ) NE forming the regular rosettes. ( l ) Developed rosette with an in-apical side. The dotted line in ( j , k ) represents the edge of the NE. White arrows in ( j, k , l ) point at the apical side. ( m ) Ratio of the developed in-apical regular rosettes in all 20% O 2 groups and 40% O 2 groups. NE: Neuroectodermal epithelium. EB: Embryonic body.
    Figure Legend Snippet: Cell proliferation within the NE requires oxygen support. ( a , c , e ) Tuj1 + Ki67, Tuj1 + NESTIN, Tuj1 + SOX2 double-stained NE in 62D 20% O 2 group. ( b , d , f ) Tuj1 + Ki67, Tuj1 + NESTIN, Tuj1 + SOX2 double-stained in 50D 40% O 2 group. White arrows in ( a , b ) point at the Ki67-positive proliferating cells in the NE of EB. The area between the two dotted lines in ( c , d ) represents the NESTIN-positive neural stem cells. The area between the two dotted lines in ( f ) represents the plate-ranged Tuj1-positive neurons. ( g , h ) Enlarged images of rosettes fusing at the edge of the EB in ( e ). ( i ) Enlarged Tuj1 (green) and SOX2 (red) double-stained images of typical rosettes fusing at the edge of EB in 50D 20% O 2 . White arrows in g , ( h , i ) point at the extroverting inner side of the rosettes. ( j – l ) ZO-1 staining of apical surface in NE. ( j ) An out-apical-side NE without forming rosettes. ( k ) NE forming the regular rosettes. ( l ) Developed rosette with an in-apical side. The dotted line in ( j , k ) represents the edge of the NE. White arrows in ( j, k , l ) point at the apical side. ( m ) Ratio of the developed in-apical regular rosettes in all 20% O 2 groups and 40% O 2 groups. NE: Neuroectodermal epithelium. EB: Embryonic body.

    Techniques Used: Staining

    Identification of H1 human embryonic stem cell (ESC) line and early embryonic body inducement. ( a – d ) Immunofluorescence of ESC markers of H1 embryonic stem cell line treated with ( a ) NANOG; ( b ) OCT4; ( c ) SOX2; ( d ) SSEA4. ( e – h ) Fluorescence-activated cell sorting analysis of the H1 embryonic stem cell line. ( e ) Negative control. ( f ) OCT4; ( g ) SOX2; ( h ) SSEA4. ( i – l ) Representative image of early stage embryonic bodies. ( i ) Day 4; ( j ) Day 8; ( k ) Day 12; ( l ) Day 16. Arrows in ( k , l ) are pointing at the neural retina at the edges of the EBs. These data revealed that the H1 embryonic stem cell line expressed the embryonic marker. Neuroectodermal epithelia were about to emerge on day 8 and were evident from day 12.
    Figure Legend Snippet: Identification of H1 human embryonic stem cell (ESC) line and early embryonic body inducement. ( a – d ) Immunofluorescence of ESC markers of H1 embryonic stem cell line treated with ( a ) NANOG; ( b ) OCT4; ( c ) SOX2; ( d ) SSEA4. ( e – h ) Fluorescence-activated cell sorting analysis of the H1 embryonic stem cell line. ( e ) Negative control. ( f ) OCT4; ( g ) SOX2; ( h ) SSEA4. ( i – l ) Representative image of early stage embryonic bodies. ( i ) Day 4; ( j ) Day 8; ( k ) Day 12; ( l ) Day 16. Arrows in ( k , l ) are pointing at the neural retina at the edges of the EBs. These data revealed that the H1 embryonic stem cell line expressed the embryonic marker. Neuroectodermal epithelia were about to emerge on day 8 and were evident from day 12.

    Techniques Used: Immunofluorescence, Fluorescence, FACS, Negative Control, Marker

    24) Product Images from "Nanog interaction with the androgen receptor signaling axis induce ovarian cancer stem cell regulation: studies based on the CRISPR/Cas9 system"

    Article Title: Nanog interaction with the androgen receptor signaling axis induce ovarian cancer stem cell regulation: studies based on the CRISPR/Cas9 system

    Journal: Journal of Ovarian Research

    doi: 10.1186/s13048-018-0403-2

    Protein expression of other stem cell genes exhibits the same tendency as Nanog. a and b ) Oct4 and Sox2 expression increased in the GFP (+) cells compared with the GFP (−) cells according to western blot analysis. c and d ) Oct4 and Sox2 expression in the GFP (+) cells clearly increased under DHT treatment compared with ASC-J9 treatment based on western blot analysis, which was also consistent with the Nanog expression trend
    Figure Legend Snippet: Protein expression of other stem cell genes exhibits the same tendency as Nanog. a and b ) Oct4 and Sox2 expression increased in the GFP (+) cells compared with the GFP (−) cells according to western blot analysis. c and d ) Oct4 and Sox2 expression in the GFP (+) cells clearly increased under DHT treatment compared with ASC-J9 treatment based on western blot analysis, which was also consistent with the Nanog expression trend

    Techniques Used: Expressing, Western Blot

    25) Product Images from "The Death-inducer Obliterator 1 (Dido1) Gene Regulates Embryonic Stem Cell Self-renewal *"

    Article Title: The Death-inducer Obliterator 1 (Dido1) Gene Regulates Embryonic Stem Cell Self-renewal *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.486290

    Dido1 feedback regulates the expression of pluripotency factors. A and B , mouse ES cells stably expressing DIDO1 were cultured in the presence of LIF and examined for the expression of Oct4 , Sox2 , and Nanog by RT-qPCR ( A ) and Western blotting ( B ). Cells
    Figure Legend Snippet: Dido1 feedback regulates the expression of pluripotency factors. A and B , mouse ES cells stably expressing DIDO1 were cultured in the presence of LIF and examined for the expression of Oct4 , Sox2 , and Nanog by RT-qPCR ( A ) and Western blotting ( B ). Cells

    Techniques Used: Expressing, Stable Transfection, Cell Culture, Quantitative RT-PCR, Western Blot

    26) Product Images from "Inhibition of H3K27me3 Histone Demethylase Activity Prevents the Proliferative Regeneration of Zebrafish Lateral Line Neuromasts"

    Article Title: Inhibition of H3K27me3 Histone Demethylase Activity Prevents the Proliferative Regeneration of Zebrafish Lateral Line Neuromasts

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2017.00051

    GSK-J4 reduced proliferation in regenerating neuromast cells. (A–D) 5 dpf larvae were treated with 400 μM neomycin for 1 h followed by GSK-J4 exposure for 24 or 48 h in the presence of BrdU. GSK-J4 significantly reduced the numbers of Sox2-positive (red) and BrdU-positive (white) replicating cells. Scale bars = 10 μm. (E,F) Quantification of Sox2-positive and BrdU-positive cells per neuromast (NM) in DMSO-treated control larvae and 10 μM GSK-J4-treated larvae at 24 or 48 h following neomycin damage. In the 24-h group, n = 30 neuromasts of DMSO-treated control larvae (15 larvae) and n = 24 neuromasts of 10 μM GSK-J4-treated larvae (12 larvae); in the 48-h group, n = 36 neuromastsof DMSO-treated control larvae (18 larvae) and n = 24 neuromasts of 10 μM GSK-J4-treated larvae (12 larvae). *** p
    Figure Legend Snippet: GSK-J4 reduced proliferation in regenerating neuromast cells. (A–D) 5 dpf larvae were treated with 400 μM neomycin for 1 h followed by GSK-J4 exposure for 24 or 48 h in the presence of BrdU. GSK-J4 significantly reduced the numbers of Sox2-positive (red) and BrdU-positive (white) replicating cells. Scale bars = 10 μm. (E,F) Quantification of Sox2-positive and BrdU-positive cells per neuromast (NM) in DMSO-treated control larvae and 10 μM GSK-J4-treated larvae at 24 or 48 h following neomycin damage. In the 24-h group, n = 30 neuromasts of DMSO-treated control larvae (15 larvae) and n = 24 neuromasts of 10 μM GSK-J4-treated larvae (12 larvae); in the 48-h group, n = 36 neuromastsof DMSO-treated control larvae (18 larvae) and n = 24 neuromasts of 10 μM GSK-J4-treated larvae (12 larvae). *** p

    Techniques Used:

    27) Product Images from "Cure of ADPKD by Selection for Spontaneous Genetic Repair Events in Pkd1-Mutated iPS Cells"

    Article Title: Cure of ADPKD by Selection for Spontaneous Genetic Repair Events in Pkd1-Mutated iPS Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0032018

    Screening of mutation-restored (Pkd1(+/R+)) iPSCs from iPSCs heterozygous for Pkd1 knockout (KO) (Pkd1(+/−)). a, Normal colony morphology of Pkd1(+/−) iPSCs expressing fluorescence marker protein, GFP. b, Expression of pluripotent marker proteins, Oct4, Sox2, and Nanog in Pkd1(+/R+) iPSCs by immunohistochemical analyses. c, Transcription of pluripotent marker genes in Pkd1(+/−) and Pkd1(+/R+) iPSCs by RT-PCR analyses. d, Secondary screening of Pkd1(+/R+) iPSCs by genomic PCR analyses. e, Verification of replacement of the KO allele by the wild-type (WT) allele through spontaneous mitotic recombination in Pkd1(+/R+) iPSCs by Southern blot hybridization analyses. Relative intensity is noted under the 15.1 kb band. f, Determination of the origin of Pkd1(+/R+) iPSCs by Southern blot hybridization.
    Figure Legend Snippet: Screening of mutation-restored (Pkd1(+/R+)) iPSCs from iPSCs heterozygous for Pkd1 knockout (KO) (Pkd1(+/−)). a, Normal colony morphology of Pkd1(+/−) iPSCs expressing fluorescence marker protein, GFP. b, Expression of pluripotent marker proteins, Oct4, Sox2, and Nanog in Pkd1(+/R+) iPSCs by immunohistochemical analyses. c, Transcription of pluripotent marker genes in Pkd1(+/−) and Pkd1(+/R+) iPSCs by RT-PCR analyses. d, Secondary screening of Pkd1(+/R+) iPSCs by genomic PCR analyses. e, Verification of replacement of the KO allele by the wild-type (WT) allele through spontaneous mitotic recombination in Pkd1(+/R+) iPSCs by Southern blot hybridization analyses. Relative intensity is noted under the 15.1 kb band. f, Determination of the origin of Pkd1(+/R+) iPSCs by Southern blot hybridization.

    Techniques Used: Mutagenesis, Knock-Out, Expressing, Fluorescence, Marker, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Southern Blot, Hybridization

    28) Product Images from "MicroRNA-194 Regulates the Development and Differentiation of Sensory Patches and Statoacoustic Ganglion of Inner Ear by Fgf4"

    Article Title: MicroRNA-194 Regulates the Development and Differentiation of Sensory Patches and Statoacoustic Ganglion of Inner Ear by Fgf4

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.906277

    MiR-194 promoted the differentiation of the sensory patches in zebrafish inner ear. The percentage of Sox2 ( upper portion ) and acetylated-tubulin ( lower portion ) positive cells were measured by in situ hybridization at 48 hpf. The white circle area indicated zebrafish inner ear. Red staining indicated Sox2 expression ( upper portion ) or acetylated-tubulin expression ( lower portion ); blue indicated cell nucleus. MO – microinjection with morpholino oligonucleotide; NC – negative control; Mimics – microinjection with miR-194 mimics; hpf – hour post fertilization.
    Figure Legend Snippet: MiR-194 promoted the differentiation of the sensory patches in zebrafish inner ear. The percentage of Sox2 ( upper portion ) and acetylated-tubulin ( lower portion ) positive cells were measured by in situ hybridization at 48 hpf. The white circle area indicated zebrafish inner ear. Red staining indicated Sox2 expression ( upper portion ) or acetylated-tubulin expression ( lower portion ); blue indicated cell nucleus. MO – microinjection with morpholino oligonucleotide; NC – negative control; Mimics – microinjection with miR-194 mimics; hpf – hour post fertilization.

    Techniques Used: In Situ Hybridization, Staining, Expressing, Negative Control

    MiR-194 promoted differentiation of SGN precursor in vitro . The proportion of Sox2 and TuJ1 positive cells were measured using immunofluorescence staining. Red staining indicated Sox2 or TuJ1 expression; Green indicated successful transfection NC, inhibitor or miR-194 mimics; Blue indicated cell nucleus. NC – negative control.
    Figure Legend Snippet: MiR-194 promoted differentiation of SGN precursor in vitro . The proportion of Sox2 and TuJ1 positive cells were measured using immunofluorescence staining. Red staining indicated Sox2 or TuJ1 expression; Green indicated successful transfection NC, inhibitor or miR-194 mimics; Blue indicated cell nucleus. NC – negative control.

    Techniques Used: In Vitro, Immunofluorescence, Staining, Expressing, Transfection, Negative Control

    MiR-194 regulated the development and differentiation of inner ear sensory patches by modulating the expression of Fgf4. The percentage of Sox2 ( left panel ) and acetylated-tubulin ( right panel ) were measured by in situ hybridization at 48 and 72 hpf. The white circle area indicated zebrafish inner ear. Red staining indicated Sox2 expression ( left panel ) or acetylated-tubulin expression ( right panel ); blue indicated cell nucleus. MO – microinjection with morpholino oligonucleotide; NC – negative control; MO+FGF4 – microinjection with morpholino oligonucleotide and Fgf4; hpf – hour post fertilization.
    Figure Legend Snippet: MiR-194 regulated the development and differentiation of inner ear sensory patches by modulating the expression of Fgf4. The percentage of Sox2 ( left panel ) and acetylated-tubulin ( right panel ) were measured by in situ hybridization at 48 and 72 hpf. The white circle area indicated zebrafish inner ear. Red staining indicated Sox2 expression ( left panel ) or acetylated-tubulin expression ( right panel ); blue indicated cell nucleus. MO – microinjection with morpholino oligonucleotide; NC – negative control; MO+FGF4 – microinjection with morpholino oligonucleotide and Fgf4; hpf – hour post fertilization.

    Techniques Used: Expressing, In Situ Hybridization, Staining, Negative Control

    29) Product Images from "Proton beam radiation induces DNA damage and cell apoptosis in glioma stem cells through reactive oxygen species"

    Article Title: Proton beam radiation induces DNA damage and cell apoptosis in glioma stem cells through reactive oxygen species

    Journal: Scientific Reports

    doi: 10.1038/srep13961

    Proton radiation induces greater cytotoxicity in GSCs than photon radiation. ( A ) T4213 CD133 + and CD133 – cells were isolated from GBM patients, and stained with anti-SOX2 and anti-GFAP antibodies and Alexa Fluor 633-phalloidin. ( B ) Schematic of the solid water phantom used for proton irradiation. Shown are where cells were contained during irradiation and where the spread-out Bragg peak fell with regards to the phantom. ( C ) IN528 GSCs were irradiated and clonogenic survival fraction determined one week after radiation (means ± SEM, n = 18 from pooled four experiments, p values determined by a Student’s t test).
    Figure Legend Snippet: Proton radiation induces greater cytotoxicity in GSCs than photon radiation. ( A ) T4213 CD133 + and CD133 – cells were isolated from GBM patients, and stained with anti-SOX2 and anti-GFAP antibodies and Alexa Fluor 633-phalloidin. ( B ) Schematic of the solid water phantom used for proton irradiation. Shown are where cells were contained during irradiation and where the spread-out Bragg peak fell with regards to the phantom. ( C ) IN528 GSCs were irradiated and clonogenic survival fraction determined one week after radiation (means ± SEM, n = 18 from pooled four experiments, p values determined by a Student’s t test).

    Techniques Used: Isolation, Staining, Irradiation

    30) Product Images from "Upregulation of SOX2-activated lncRNA ANRIL promotes nasopharyngeal carcinoma cell growth"

    Article Title: Upregulation of SOX2-activated lncRNA ANRIL promotes nasopharyngeal carcinoma cell growth

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21708-z

    SOX2 induces the expression of ANRIL by promoting its transcription. ( A ) Bioinformatics analysis of the binding site of SOX2 on the ANRIL promoter. ChIP-qPCR ( B ) and dual luciferase assay ( C ) were used to confirm the binding of SOX2 with the ANRIL promoter. ( D ) Quantitative real-time RT-PCR analysed the correlation of ANRIL and SOX2 transcripts. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: SOX2 induces the expression of ANRIL by promoting its transcription. ( A ) Bioinformatics analysis of the binding site of SOX2 on the ANRIL promoter. ChIP-qPCR ( B ) and dual luciferase assay ( C ) were used to confirm the binding of SOX2 with the ANRIL promoter. ( D ) Quantitative real-time RT-PCR analysed the correlation of ANRIL and SOX2 transcripts. Data are presented as the mean ± SD. *P

    Techniques Used: Expressing, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Luciferase, Quantitative RT-PCR

    ANRIL is required for SOX2-driven nasopharyngeal carcinoma proliferation. ( A ) ANRIL expression levels in HNE-1 and CNE2 cells transfected with SOX2 shRNAs alone or in combination with the ANRIL vector were analysed by quantitative real-time RT-PCR assays. Cell growth ( B ) and colony formation ( C ) were analysed in HNE-1 and CNE2 with SOX2 shRNAs alone or in combination with ANRIL. Data are presented as the mean ± SD. *P
    Figure Legend Snippet: ANRIL is required for SOX2-driven nasopharyngeal carcinoma proliferation. ( A ) ANRIL expression levels in HNE-1 and CNE2 cells transfected with SOX2 shRNAs alone or in combination with the ANRIL vector were analysed by quantitative real-time RT-PCR assays. Cell growth ( B ) and colony formation ( C ) were analysed in HNE-1 and CNE2 with SOX2 shRNAs alone or in combination with ANRIL. Data are presented as the mean ± SD. *P

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR

    ANRIL/β-catenin is essential for SOX2-mediated nasopharyngeal carcinoma progression. ( A ) Levels of SOX2 and β-catenin protein expression in HNE-1 and CNE2 after stably sh-SOX2 and shControl. ( B ) β-catenin protein expression analysis in HNE-1 and CNE2 with SOX2 shRNAs alone or in combination with ANRIL vector. ( C ) Schematic diagram indicated the β-catenin binding to ANRIL region by RNA IP assays. ( D ) TOP-FOP flash assay was performed to evaluate the effect of shANRIL on WNT/β-catenin pathway. ( E ) Correlation between ANRIL and β-catenin transcript determined by quantitative real-time RT-PCR in 20 human nasopharyngeal carcinoma specimens.
    Figure Legend Snippet: ANRIL/β-catenin is essential for SOX2-mediated nasopharyngeal carcinoma progression. ( A ) Levels of SOX2 and β-catenin protein expression in HNE-1 and CNE2 after stably sh-SOX2 and shControl. ( B ) β-catenin protein expression analysis in HNE-1 and CNE2 with SOX2 shRNAs alone or in combination with ANRIL vector. ( C ) Schematic diagram indicated the β-catenin binding to ANRIL region by RNA IP assays. ( D ) TOP-FOP flash assay was performed to evaluate the effect of shANRIL on WNT/β-catenin pathway. ( E ) Correlation between ANRIL and β-catenin transcript determined by quantitative real-time RT-PCR in 20 human nasopharyngeal carcinoma specimens.

    Techniques Used: Expressing, Stable Transfection, Plasmid Preparation, Binding Assay, Quantitative RT-PCR

    31) Product Images from "Position-dependent plasticity of distinct progenitor types in the primitive streak"

    Article Title: Position-dependent plasticity of distinct progenitor types in the primitive streak

    Journal: eLife

    doi: 10.7554/eLife.10042

    Homotopic and heterotopic grafts incorporate well into host embryos. ( A – C ) Immunohistochemical confirmation of differentiation markers in different graft series. Upper row, DAPI-counterstained images (grey). Boxes, region magnified in lower row. Lower row, immunofluorescence channel. Position of grafted donor cells is outlined in white. ( A ) Sox2, T, and Foxa2 expression in axis and TB sections of L1 homotopic grafts. Note lack of notochord-specific Foxa2 expression in graft-derived cells. ( B ) Sox2 and T expression in axis and TB sections of L2-3 heterotopic grafts (to Br). ( C ) PDGFRβ expression in L/St5-derived cells, grafted either homo- or heterotopically to a wildtype host. ( D ) Table showing the number of embryos and sections stained for each marker. ( E ) Overview of marker expression in different regions of the axis. Note that none of the CLE-derived grafted cells expressed FoxA2 or high levels of T protein in the caudal notochord domain. DOI: http://dx.doi.org/10.7554/eLife.10042.014
    Figure Legend Snippet: Homotopic and heterotopic grafts incorporate well into host embryos. ( A – C ) Immunohistochemical confirmation of differentiation markers in different graft series. Upper row, DAPI-counterstained images (grey). Boxes, region magnified in lower row. Lower row, immunofluorescence channel. Position of grafted donor cells is outlined in white. ( A ) Sox2, T, and Foxa2 expression in axis and TB sections of L1 homotopic grafts. Note lack of notochord-specific Foxa2 expression in graft-derived cells. ( B ) Sox2 and T expression in axis and TB sections of L2-3 heterotopic grafts (to Br). ( C ) PDGFRβ expression in L/St5-derived cells, grafted either homo- or heterotopically to a wildtype host. ( D ) Table showing the number of embryos and sections stained for each marker. ( E ) Overview of marker expression in different regions of the axis. Note that none of the CLE-derived grafted cells expressed FoxA2 or high levels of T protein in the caudal notochord domain. DOI: http://dx.doi.org/10.7554/eLife.10042.014

    Techniques Used: Immunohistochemistry, Immunofluorescence, Expressing, Derivative Assay, Staining, Marker

    Rostral CLE tissue contains Sox2 + T + cells. Graft donor tissue was examined for the presence of Sox2/T coexpressing cells. L1-3 and L/St5 pieces were dissected as described above. The underlying presomitic mesoderm was manually removed in the L1-3 grafts. ( A ) L1-3 donor tissue contains Sox2 + T + cells (n = 7/7). ( B ) L/St5 were devoid of coexpressing cells (n = 5/5). Blue, DAPI nuclear stain; green, Sox2; red, T. DOI: http://dx.doi.org/10.7554/eLife.10042.012
    Figure Legend Snippet: Rostral CLE tissue contains Sox2 + T + cells. Graft donor tissue was examined for the presence of Sox2/T coexpressing cells. L1-3 and L/St5 pieces were dissected as described above. The underlying presomitic mesoderm was manually removed in the L1-3 grafts. ( A ) L1-3 donor tissue contains Sox2 + T + cells (n = 7/7). ( B ) L/St5 were devoid of coexpressing cells (n = 5/5). Blue, DAPI nuclear stain; green, Sox2; red, T. DOI: http://dx.doi.org/10.7554/eLife.10042.012

    Techniques Used: Staining

    NMPs express low levels of T and Sox2. ( A–C ) 3D analysis showing the relative levels of Sox2 and T protein in the E8.5–10.5 caudal region. Sox2 + T + cells express low-to-medium levels of both transcription factors (green cells in Ac, Ad, Bc and Cc represent Sox2 + T + cells, indicated by the area within the white dotted line). Colours represent the intensity range shown as arbitrary units (AU). The lower threshold for positivity was calculated as before (see Figure 3—figure supplement 1B ), with the maximum corresponding to the highest intensity recorded in the z-stack. Asterisk, Sox2 + T + cells found in the dorsocaudal part of the gut; grey, segmented nuclear volumes negative for either transcription factor. DOI: http://dx.doi.org/10.7554/eLife.10042.009
    Figure Legend Snippet: NMPs express low levels of T and Sox2. ( A–C ) 3D analysis showing the relative levels of Sox2 and T protein in the E8.5–10.5 caudal region. Sox2 + T + cells express low-to-medium levels of both transcription factors (green cells in Ac, Ad, Bc and Cc represent Sox2 + T + cells, indicated by the area within the white dotted line). Colours represent the intensity range shown as arbitrary units (AU). The lower threshold for positivity was calculated as before (see Figure 3—figure supplement 1B ), with the maximum corresponding to the highest intensity recorded in the z-stack. Asterisk, Sox2 + T + cells found in the dorsocaudal part of the gut; grey, segmented nuclear volumes negative for either transcription factor. DOI: http://dx.doi.org/10.7554/eLife.10042.009

    Techniques Used:

    Grafts using β-catenin knock out donor tissue. ( A – C ) Grafting scheme of L1-3 ( A ) or L/St5 ( B ) to St1-3 heterotopic grafts. Either AGFP7 control or βcatCKO:sGFP donor tissue was used to transplant into WT host embryos. ( C ) After receiving the graft, embryos were grown in vitro for 48 hr, of which the first 8 hr in the presence of 10 µM 4-OHT. ( D ) Diagram summarising experimental outcome for L1-3 or L/St5 to St1-3 heterotopic grafts. ( E ) Number of embryo grafts performed. ( F ) Exceptional L1-3 AGFP7 to St1-3 grafted embryo showing minor neural CNH contribution, shown by Sox2 immunostaining in the tail bud. ( G ) DAPI-counterstained images (grey). Immunofluorescent staining for neural Sox2, FoxA2 and Pax6 differentiation, and dermomyotome-specific Pax3 after 48 hr in vitro culture. ( H ) Example of a L/St5 βcatCKO:sGFP to St1-3 graft that contained non-integrated clumps (arrowhead) near the notochord (n = 3/7). These non-integrated cell clumps did not express Sox2, T or FoxA2. DOI: http://dx.doi.org/10.7554/eLife.10042.020
    Figure Legend Snippet: Grafts using β-catenin knock out donor tissue. ( A – C ) Grafting scheme of L1-3 ( A ) or L/St5 ( B ) to St1-3 heterotopic grafts. Either AGFP7 control or βcatCKO:sGFP donor tissue was used to transplant into WT host embryos. ( C ) After receiving the graft, embryos were grown in vitro for 48 hr, of which the first 8 hr in the presence of 10 µM 4-OHT. ( D ) Diagram summarising experimental outcome for L1-3 or L/St5 to St1-3 heterotopic grafts. ( E ) Number of embryo grafts performed. ( F ) Exceptional L1-3 AGFP7 to St1-3 grafted embryo showing minor neural CNH contribution, shown by Sox2 immunostaining in the tail bud. ( G ) DAPI-counterstained images (grey). Immunofluorescent staining for neural Sox2, FoxA2 and Pax6 differentiation, and dermomyotome-specific Pax3 after 48 hr in vitro culture. ( H ) Example of a L/St5 βcatCKO:sGFP to St1-3 graft that contained non-integrated clumps (arrowhead) near the notochord (n = 3/7). These non-integrated cell clumps did not express Sox2, T or FoxA2. DOI: http://dx.doi.org/10.7554/eLife.10042.020

    Techniques Used: Knock-Out, In Vitro, Immunostaining, Staining

    Sox2 + T + NMPs peak at mid trunk formation. ( A ) 3D reconstruction of the E8.5 caudal region. ( Aa–b ) Dorsal view. ( Ac ) Frontal view along the PS and CLE. Colours show different thresholded Sox2/T populations. ( B–C ) Quantification of different Sox2/T populations in the caudal embryo shows a peak in overall cell labelling at E10.5 ( B ). Sox2 + T + cell numbers are highest at E9.5 ( C ). Data in graphs is shown as the mean ± s.d. See also Figure 3—figure supplement 1B–E and Video 1 . DOI: http://dx.doi.org/10.7554/eLife.10042.007
    Figure Legend Snippet: Sox2 + T + NMPs peak at mid trunk formation. ( A ) 3D reconstruction of the E8.5 caudal region. ( Aa–b ) Dorsal view. ( Ac ) Frontal view along the PS and CLE. Colours show different thresholded Sox2/T populations. ( B–C ) Quantification of different Sox2/T populations in the caudal embryo shows a peak in overall cell labelling at E10.5 ( B ). Sox2 + T + cell numbers are highest at E9.5 ( C ). Data in graphs is shown as the mean ± s.d. See also Figure 3—figure supplement 1B–E and Video 1 . DOI: http://dx.doi.org/10.7554/eLife.10042.007

    Techniques Used:

    Quantifying Sox2 + T + cells during axis elongation. ( A ) Double in situ hybridisation for Sox2 and T at the end of axis elongation (E13.5, 65s). ( B ) Workflow for Sox2/T quantification and 3D reconstruction. ( C and F ) Total cells analysed per embryonic stage ( C ) or in 4-OHT-treated βcatCKO embryos ( F ). Data is shown as the mean ± s.d. ( D and G ) Quantification of Sox2/T populations per embryonic stage ( D ) or in 4-OHT-treated βcatCKO embryos ( G ). Data is shown as the mean (s.d.) with n, the number of embryos analysed. ( E and H ) Statistical analysis. ( E ) A standard, unpaired Student’s t-test was used to test the significance in Sox2 + T + cell number and the total cells analysed between consecutive stages. ( H ) P-values obtained through an unpaired Student's t-test (with Welch’s correction) comparing Sox2/T populations in E9.5 and E8.5 wildtype to E9.5 4-OHT-treated βcatCKO embryos (*, p-value
    Figure Legend Snippet: Quantifying Sox2 + T + cells during axis elongation. ( A ) Double in situ hybridisation for Sox2 and T at the end of axis elongation (E13.5, 65s). ( B ) Workflow for Sox2/T quantification and 3D reconstruction. ( C and F ) Total cells analysed per embryonic stage ( C ) or in 4-OHT-treated βcatCKO embryos ( F ). Data is shown as the mean ± s.d. ( D and G ) Quantification of Sox2/T populations per embryonic stage ( D ) or in 4-OHT-treated βcatCKO embryos ( G ). Data is shown as the mean (s.d.) with n, the number of embryos analysed. ( E and H ) Statistical analysis. ( E ) A standard, unpaired Student’s t-test was used to test the significance in Sox2 + T + cell number and the total cells analysed between consecutive stages. ( H ) P-values obtained through an unpaired Student's t-test (with Welch’s correction) comparing Sox2/T populations in E9.5 and E8.5 wildtype to E9.5 4-OHT-treated βcatCKO embryos (*, p-value

    Techniques Used: In Situ, Hybridization

    Position-dependent plasticity of primitive streak progenitors. ( A ) NMPs and LPMPs coexist in the caudal progenitor zone. The NSB and L1 regions show NM fate. NM potency is broader, encompassing L2–3 regions and coincides with a Sox2 + T + phenotype. LPMPs reside in the caudal-most embryo part; their potency to make PXM is somehow supressed. ( B ) Relative N vs M fate choices and trajectories for both progenitor populations are represented by directional purple and cyan arrows respectively, with length indicating proportions of cells entering each lineage. Background, relative expression of Sox2 (green) and T (red). Only NMPs require Wnt/β-catenin for mesoderm differentiation and maintenance. DOI: http://dx.doi.org/10.7554/eLife.10042.026
    Figure Legend Snippet: Position-dependent plasticity of primitive streak progenitors. ( A ) NMPs and LPMPs coexist in the caudal progenitor zone. The NSB and L1 regions show NM fate. NM potency is broader, encompassing L2–3 regions and coincides with a Sox2 + T + phenotype. LPMPs reside in the caudal-most embryo part; their potency to make PXM is somehow supressed. ( B ) Relative N vs M fate choices and trajectories for both progenitor populations are represented by directional purple and cyan arrows respectively, with length indicating proportions of cells entering each lineage. Background, relative expression of Sox2 (green) and T (red). Only NMPs require Wnt/β-catenin for mesoderm differentiation and maintenance. DOI: http://dx.doi.org/10.7554/eLife.10042.026

    Techniques Used: Expressing

    32) Product Images from "Directed reprogramming of comprehensively characterized dental pulp stem cells extracted from natal tooth"

    Article Title: Directed reprogramming of comprehensively characterized dental pulp stem cells extracted from natal tooth

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24421-z

    ( a ) nDPSC derived hiPSC. Image of nDPSC derived hiPSC with typical hES like morphology. ( b ) Colorimetric detection of alkaline phosphatase. ( c – f ) Immunocytochemistry against ( c ) SSEA-4, ( d ) POU5F1, ( e ) SOX2, and ( f ) NANOG. Nuclei were counterstained with DAPI. Images are shown as overlap of the two channels ( c – f ). Scale bar = 200 µm.
    Figure Legend Snippet: ( a ) nDPSC derived hiPSC. Image of nDPSC derived hiPSC with typical hES like morphology. ( b ) Colorimetric detection of alkaline phosphatase. ( c – f ) Immunocytochemistry against ( c ) SSEA-4, ( d ) POU5F1, ( e ) SOX2, and ( f ) NANOG. Nuclei were counterstained with DAPI. Images are shown as overlap of the two channels ( c – f ). Scale bar = 200 µm.

    Techniques Used: Derivative Assay, Immunocytochemistry

    33) Product Images from "Tracking diphyodont development in miniature pigs in vitro and in vivo"

    Article Title: Tracking diphyodont development in miniature pigs in vitro and in vivo

    Journal: Biology Open

    doi: 10.1242/bio.037036

    The expression of candidate genes in the developing mandibular p4 and successional dental lamina in frontal sections of WZSP embryo. (A) SOX2, (B) BMP4 and (C) WNT10b immunohistochemical stain. (A–C) Right panels show the enlarged successional dental lamina, shown with an arrowhead inside the boxed area in the left panels. Scale bars: 500 μm (left panels), 50 μm (right panels).
    Figure Legend Snippet: The expression of candidate genes in the developing mandibular p4 and successional dental lamina in frontal sections of WZSP embryo. (A) SOX2, (B) BMP4 and (C) WNT10b immunohistochemical stain. (A–C) Right panels show the enlarged successional dental lamina, shown with an arrowhead inside the boxed area in the left panels. Scale bars: 500 μm (left panels), 50 μm (right panels).

    Techniques Used: Expressing, Immunohistochemistry, Staining

    34) Product Images from "LncRNA ADAMTS9‐AS2 suppresses the proliferation of gastric cancer cells and the tumorigenicity of cancer stem cells through regulating SPOP, et al. LncRNA ADAMTS9‐AS2 suppresses the proliferation of gastric cancer cells and the tumorigenicity of cancer stem cells through regulating SPOP"

    Article Title: LncRNA ADAMTS9‐AS2 suppresses the proliferation of gastric cancer cells and the tumorigenicity of cancer stem cells through regulating SPOP, et al. LncRNA ADAMTS9‐AS2 suppresses the proliferation of gastric cancer cells and the tumorigenicity of cancer stem cells through regulating SPOP

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.15161

    SPOP regulates stem‐like capacities of gastric CSC. A,B, The mRNA and protein expression levels of CSCs markers (CD44, Oct4, Sox2 and NANOG) were examined by qPCR and Western blot in tumorsphere cells SPOP overexpression plasmid and control vector cells. C,D, The mRNA and protein expression levels of CD44, Oct4, Sox2 and NANOG in tumorsphere cells transfected with SPOP siRNA or scramble siRNA were also examined by qPCR and Western blot analysis. E. A number of spheres per well were quantified on spheroid formation assay for tumorsphere cells transfected with SPOP overexpression plasmid or SPOP siRNA, respectively. * P
    Figure Legend Snippet: SPOP regulates stem‐like capacities of gastric CSC. A,B, The mRNA and protein expression levels of CSCs markers (CD44, Oct4, Sox2 and NANOG) were examined by qPCR and Western blot in tumorsphere cells SPOP overexpression plasmid and control vector cells. C,D, The mRNA and protein expression levels of CD44, Oct4, Sox2 and NANOG in tumorsphere cells transfected with SPOP siRNA or scramble siRNA were also examined by qPCR and Western blot analysis. E. A number of spheres per well were quantified on spheroid formation assay for tumorsphere cells transfected with SPOP overexpression plasmid or SPOP siRNA, respectively. * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Plasmid Preparation, Transfection, Tube Formation Assay

    MKN‐45 cells formed the anchorage‐independent, self‐renewing spheroid bodies. A, The generation of a spheroid body from a single MKN‐45 cell was cultured in a 96‐well dish. Observation time‐point: days 0, 3, 7, 10, 14 and 21 and photographed under the light microscope (magnification × 200). B, Western blot analysis was used on adherent cells and tumorsphere cells, which indicated that the expression levels of Oct3/4, Sox2 and CD44 were dramatically more in tumorsphere cells than in adherent cells. C, Intracellular localization of Oct3/4, Sox2 and CD44 in tumorsphere cells by using immunofluorescence staining. And dual staining of Oct3/4 and Sox2 showed that Oct3/4‐positive stained cells were co‐stained with Sox2. DAPI was applied for the nuclear counterstain
    Figure Legend Snippet: MKN‐45 cells formed the anchorage‐independent, self‐renewing spheroid bodies. A, The generation of a spheroid body from a single MKN‐45 cell was cultured in a 96‐well dish. Observation time‐point: days 0, 3, 7, 10, 14 and 21 and photographed under the light microscope (magnification × 200). B, Western blot analysis was used on adherent cells and tumorsphere cells, which indicated that the expression levels of Oct3/4, Sox2 and CD44 were dramatically more in tumorsphere cells than in adherent cells. C, Intracellular localization of Oct3/4, Sox2 and CD44 in tumorsphere cells by using immunofluorescence staining. And dual staining of Oct3/4 and Sox2 showed that Oct3/4‐positive stained cells were co‐stained with Sox2. DAPI was applied for the nuclear counterstain

    Techniques Used: Cell Culture, Light Microscopy, Western Blot, Expressing, Immunofluorescence, Staining

    35) Product Images from "Core transcription factors, Oct4, Sox2 and Nanog, individually form complexes with nucleophosmin (Npm1) to control embryonic stem (ES) cell fate determination"

    Article Title: Core transcription factors, Oct4, Sox2 and Nanog, individually form complexes with nucleophosmin (Npm1) to control embryonic stem (ES) cell fate determination

    Journal: Aging (Albany NY)

    doi:

    Sox2 physically interacts with Npm1 in ES cells. ( A ) Immunofluorescence confocal microscopy in combination with in situ PLA showed that there is an interaction between Sox2 and Npm1 in ES cells. Complexes (red dots) were detected in the nucleoplasm of interphase cells. DNA was counterstained by Hoechst 33342 (blue). Scale bar represents 10 μm. ( B ) Co-immunoprecipitation experiments followed by Western blot analysis showed that Npm1 can be immunoprecipitated using anti-Sox2 (1 M NaCl and 0.1 M Citrate).
    Figure Legend Snippet: Sox2 physically interacts with Npm1 in ES cells. ( A ) Immunofluorescence confocal microscopy in combination with in situ PLA showed that there is an interaction between Sox2 and Npm1 in ES cells. Complexes (red dots) were detected in the nucleoplasm of interphase cells. DNA was counterstained by Hoechst 33342 (blue). Scale bar represents 10 μm. ( B ) Co-immunoprecipitation experiments followed by Western blot analysis showed that Npm1 can be immunoprecipitated using anti-Sox2 (1 M NaCl and 0.1 M Citrate).

    Techniques Used: Immunofluorescence, Confocal Microscopy, In Situ, Proximity Ligation Assay, Immunoprecipitation, Western Blot

    Tpt1 and Npm1 are involved in ES cell maintenance. Involvement of Tpt1 and Npm1 in ES cell maintenance was investigated using shRNA constructs against each gene and analyzed by qPCR 48 h post transfection. Downregulation of Tpt1 (blue bars) resulted in a minor increase in Oct4 and Brachyury (Bra) levels, a decrease in GATA4 levels, whereas levels of Sox2 and Nestin (Nes) increased significantly, indicative of involvement in ectodermal differentiation. Downregulation of Npm1 (black bars) resulted in minor increases in Oct4 and Sox2 levels, decreased GATA4 levels, while increased Brachyury (Bra) levels, indicative of involvement in mesodermal differentiation. All experiments were done in triplicates and normalized first to GAPDH (reference gene), and then to corresponding negative control shRNA constructs.
    Figure Legend Snippet: Tpt1 and Npm1 are involved in ES cell maintenance. Involvement of Tpt1 and Npm1 in ES cell maintenance was investigated using shRNA constructs against each gene and analyzed by qPCR 48 h post transfection. Downregulation of Tpt1 (blue bars) resulted in a minor increase in Oct4 and Brachyury (Bra) levels, a decrease in GATA4 levels, whereas levels of Sox2 and Nestin (Nes) increased significantly, indicative of involvement in ectodermal differentiation. Downregulation of Npm1 (black bars) resulted in minor increases in Oct4 and Sox2 levels, decreased GATA4 levels, while increased Brachyury (Bra) levels, indicative of involvement in mesodermal differentiation. All experiments were done in triplicates and normalized first to GAPDH (reference gene), and then to corresponding negative control shRNA constructs.

    Techniques Used: shRNA, Construct, Real-time Polymerase Chain Reaction, Transfection, Negative Control

    Npm1/Sox2 interaction changes during differentiation. Induced differentiation of ES cells by withdrawal of leukemia inhibitory factor (LIF), addition of either dimethyl sulfoxide (DMSO) or retinoic acid (RA) for 24 h in combination with in situ PLA, showed that the Npm1/Sox2 complexes decreased during DMSO and leukemia inhibitory factor withdrawal induced differentiation. In contrast, retinoic acid induced differentiation did not notably affect the number of Npm1/Sox2 complexes in the nucleoplasm. DNA was counterstained with Hoechst 33342 (blue). Scale bar represents 10 μm.
    Figure Legend Snippet: Npm1/Sox2 interaction changes during differentiation. Induced differentiation of ES cells by withdrawal of leukemia inhibitory factor (LIF), addition of either dimethyl sulfoxide (DMSO) or retinoic acid (RA) for 24 h in combination with in situ PLA, showed that the Npm1/Sox2 complexes decreased during DMSO and leukemia inhibitory factor withdrawal induced differentiation. In contrast, retinoic acid induced differentiation did not notably affect the number of Npm1/Sox2 complexes in the nucleoplasm. DNA was counterstained with Hoechst 33342 (blue). Scale bar represents 10 μm.

    Techniques Used: In Situ, Proximity Ligation Assay

    36) Product Images from "Identification and characterisation of NANOG+/ OCT-4high/SOX2+ doxorubicin-resistant stem-like cells from transformed trophoblastic cell lines"

    Article Title: Identification and characterisation of NANOG+/ OCT-4high/SOX2+ doxorubicin-resistant stem-like cells from transformed trophoblastic cell lines

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24151

    Expression of NANOG, OCT-4 and SOX2 in transformed trophoblast spheroids and parental cells (A) (B) Immunofluorescence micrographs of OCT-4, SOX2 and NANOG with DAPI staining in HTR8/Svneo and TEV-1 transformed parental trophoblast cell lines. (B) Immunofluorescence micrographs OCT-4, SOX2 and NANOG with DAPI staining in transformed trophoblast spheroids. Objective magnification 40X. Scale bar = 50μm.
    Figure Legend Snippet: Expression of NANOG, OCT-4 and SOX2 in transformed trophoblast spheroids and parental cells (A) (B) Immunofluorescence micrographs of OCT-4, SOX2 and NANOG with DAPI staining in HTR8/Svneo and TEV-1 transformed parental trophoblast cell lines. (B) Immunofluorescence micrographs OCT-4, SOX2 and NANOG with DAPI staining in transformed trophoblast spheroids. Objective magnification 40X. Scale bar = 50μm.

    Techniques Used: Expressing, Transformation Assay, Immunofluorescence, Staining

    OCT-4, SOX2 and NANOG expression in untreated and treated transformed parental trophoblast cells and spheroids (A) (B) (C) (D) Immunoblots showing the expression of OCT-4, SOX2, NANOG, CDX2 and corresponding β-actin loading controls in HTR8/SVneo and TEV-1 untreated and treated transformed parental trophoblast cells and spheroids.
    Figure Legend Snippet: OCT-4, SOX2 and NANOG expression in untreated and treated transformed parental trophoblast cells and spheroids (A) (B) (C) (D) Immunoblots showing the expression of OCT-4, SOX2, NANOG, CDX2 and corresponding β-actin loading controls in HTR8/SVneo and TEV-1 untreated and treated transformed parental trophoblast cells and spheroids.

    Techniques Used: Expressing, Transformation Assay, Western Blot

    37) Product Images from "Single-cell analysis supports a luminal-neuroendocrine trans-differentiation in human prostate cancer"

    Article Title: Single-cell analysis supports a luminal-neuroendocrine trans-differentiation in human prostate cancer

    Journal: bioRxiv

    doi: 10.1101/2020.05.12.091132

    Intratumor heterogeneity analyses reveal different extents of NE differentiation ( A ) UMAP visualization of epithelial cell sub-clusters from each sample. ( B ) Heatmap depicting prostate lineage marker genes and AR pathway gene expression levels in epithelial cell sub-clusters from each sample. Those high-lighted in yellow frame showed cluster 5 in patient #4 and cluster 4 in patient #6 was NE sub-clusters. ( C ) Immunohistochemistry (IHC) staining for AR, SYP and SOX2 in sections from 5 samples. Scale bars represent 50μm.
    Figure Legend Snippet: Intratumor heterogeneity analyses reveal different extents of NE differentiation ( A ) UMAP visualization of epithelial cell sub-clusters from each sample. ( B ) Heatmap depicting prostate lineage marker genes and AR pathway gene expression levels in epithelial cell sub-clusters from each sample. Those high-lighted in yellow frame showed cluster 5 in patient #4 and cluster 4 in patient #6 was NE sub-clusters. ( C ) Immunohistochemistry (IHC) staining for AR, SYP and SOX2 in sections from 5 samples. Scale bars represent 50μm.

    Techniques Used: Marker, Expressing, Immunohistochemistry, Staining

    38) Product Images from "Combined Transplantation of Olfactory Ensheathing Cells With Rat Neural Stem Cells Enhanced the Therapeutic Effect in the Retina of RCS Rats"

    Article Title: Combined Transplantation of Olfactory Ensheathing Cells With Rat Neural Stem Cells Enhanced the Therapeutic Effect in the Retina of RCS Rats

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2020.00052

    Activation of endogenous retinal stem cells after combined transplantation of rat olfactory ensheathing cells (OECs) and neural stem cells (NSCs) into the subretinal space of Royal College of Surgeons (RCS) rats. (A–E) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 4 weeks postoperation. (F–J) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 8 weeks postoperation. (K–O) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 12 weeks postoperation. (P) Statistical analysis of the numbers of Sox2-positive cells in transplantation groups. Scale bar: (A–O) 50 μm. ∗ P
    Figure Legend Snippet: Activation of endogenous retinal stem cells after combined transplantation of rat olfactory ensheathing cells (OECs) and neural stem cells (NSCs) into the subretinal space of Royal College of Surgeons (RCS) rats. (A–E) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 4 weeks postoperation. (F–J) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 8 weeks postoperation. (K–O) Immunofluorescence of Sox2 in the blank, PBS, OEC, NSC, and OEC + NSC groups at 12 weeks postoperation. (P) Statistical analysis of the numbers of Sox2-positive cells in transplantation groups. Scale bar: (A–O) 50 μm. ∗ P

    Techniques Used: Activation Assay, Transplantation Assay, Immunofluorescence

    The influence on differentiation of neural stem cells (NSCs) after in vitro coculture with olfactory ensheathing cells (OECs) via Transwell system. (A–H) Immunofluorescence analysis of BrdU, Pax6, Sox2, and glial fibrillary acidic protein (GFAP) in NSCs cultured along group (A–D) and NSCs + OECs coculture group (E–H) . (I) The schematic diagram of the differentiation assay performed via Transwell system. (J) Relative statistical analysis of (A–H) . Scale bar: (A–H) 25 μm. *** P
    Figure Legend Snippet: The influence on differentiation of neural stem cells (NSCs) after in vitro coculture with olfactory ensheathing cells (OECs) via Transwell system. (A–H) Immunofluorescence analysis of BrdU, Pax6, Sox2, and glial fibrillary acidic protein (GFAP) in NSCs cultured along group (A–D) and NSCs + OECs coculture group (E–H) . (I) The schematic diagram of the differentiation assay performed via Transwell system. (J) Relative statistical analysis of (A–H) . Scale bar: (A–H) 25 μm. *** P

    Techniques Used: In Vitro, Immunofluorescence, Cell Culture, Differentiation Assay

    39) Product Images from "Transplantation of human induced cerebellar granular-like cells improves motor functions in a novel mouse model of cerebellar ataxia"

    Article Title: Transplantation of human induced cerebellar granular-like cells improves motor functions in a novel mouse model of cerebellar ataxia

    Journal: American Journal of Translational Research

    doi:

    Induction of human induced cerebellar granular-like cells (hiCGCs) from patient-derived fibroblasts. A. Ascl1, Ngn2, and Sox2 transduced fibroblasts exhibited a mature granular morphology resembling neurons at 21 days after transduction. B. Quantification of GFP/Tuj1 double-positive hiCGCs. C. The percentage of Tuj1/GFP and Ath1/Tuj1 double-positive hiCGCs reflects the hiCGCs conversion efficiency. D. Immunocytochemistry of Tuj1, Map2 and Synapsin.
    Figure Legend Snippet: Induction of human induced cerebellar granular-like cells (hiCGCs) from patient-derived fibroblasts. A. Ascl1, Ngn2, and Sox2 transduced fibroblasts exhibited a mature granular morphology resembling neurons at 21 days after transduction. B. Quantification of GFP/Tuj1 double-positive hiCGCs. C. The percentage of Tuj1/GFP and Ath1/Tuj1 double-positive hiCGCs reflects the hiCGCs conversion efficiency. D. Immunocytochemistry of Tuj1, Map2 and Synapsin.

    Techniques Used: Derivative Assay, Transduction, Immunocytochemistry

    40) Product Images from "JNK Inhibition Inhibits Lateral Line Neuromast Hair Cell Development"

    Article Title: JNK Inhibition Inhibits Lateral Line Neuromast Hair Cell Development

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2016.00019

    Detection of neuromast hair cells and supporting cells in 5 dpf larvae . (A,B) Confocal images of neuromasts from a 5 dpf control larva and 5 dpf larva treated with 10 μM SP600125. The neuromast hair cells in the transgenic line Brn3c: mGFP were detected by GFP visualization (green), supporting cells were detected by Sox2 (red), and proliferating cells were detected by BrdU (white). Higher magnification of hair cells and supporting cells of the neuromast taken from z-stacks show that hair cells and supporting cells in untreated controls and SP600125-treated animals had no observable morphological differences though there were fewer GFP-positive and Sox2-positive cells in the neuromasts of larvae treated with SP600125. The number of BrdU-labeled cells is much larger in control than in SP600125-treated larvae. Scale bar = 10 μm. (C,D) Quantification of hair cells and supporting cells in the neuromast (NM) for each experimental condition. (E–G) Quantification of replicating cells in the neuromasts for each experimental condition. SP600125 treatment decreased the numbers of BrdU-positive cells, the ratio of BrdU-positive hair cells, and the ratio of BrdU-positive supporting cells in neuromasts. The first four neuromasts along the body, L1–L4, were recorded on one side of each fish [one-way ANOVA; GFP+ cells: F (3, 112) = 237.5, p
    Figure Legend Snippet: Detection of neuromast hair cells and supporting cells in 5 dpf larvae . (A,B) Confocal images of neuromasts from a 5 dpf control larva and 5 dpf larva treated with 10 μM SP600125. The neuromast hair cells in the transgenic line Brn3c: mGFP were detected by GFP visualization (green), supporting cells were detected by Sox2 (red), and proliferating cells were detected by BrdU (white). Higher magnification of hair cells and supporting cells of the neuromast taken from z-stacks show that hair cells and supporting cells in untreated controls and SP600125-treated animals had no observable morphological differences though there were fewer GFP-positive and Sox2-positive cells in the neuromasts of larvae treated with SP600125. The number of BrdU-labeled cells is much larger in control than in SP600125-treated larvae. Scale bar = 10 μm. (C,D) Quantification of hair cells and supporting cells in the neuromast (NM) for each experimental condition. (E–G) Quantification of replicating cells in the neuromasts for each experimental condition. SP600125 treatment decreased the numbers of BrdU-positive cells, the ratio of BrdU-positive hair cells, and the ratio of BrdU-positive supporting cells in neuromasts. The first four neuromasts along the body, L1–L4, were recorded on one side of each fish [one-way ANOVA; GFP+ cells: F (3, 112) = 237.5, p

    Techniques Used: Transgenic Assay, Labeling, Fluorescence In Situ Hybridization

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    Article Snippet: .. Following heating at 100°C for 10 min in the presence of a loading buffer, equal amounts of protein lysates (50 µg) were separated using 10% SDS-PAGE (Bio-West Inc., Logan, UT, USA) at 100 V for 1 h, and transferred onto Invitrogen nitrocellulose membranes (Thermo Fisher Scientific, Inc.) at 120 V for 1 h. Following blocking with 5% skimmed milk (diluted with PBS), the membranes were incubated overnight at 4°C with the following primary antibodies: CYP27A1 (ab126785; 1:500; Abcam, Cambridge, MA, USA), c-myc (ab32072; 1:500; Abcam), RB (ab181616; 1:500; Abcam), Ki-67 (ab15580; 1:500; Abcam), CDK2 (ab32147; 1:500; Abcam), p21 (ab109520; 1:500; Abcam), p53 (ab32049; 1:500; Abcam), PDCD4 (ab51495; 1:500; Abcam), SOX2 (ab92494; 1:500; Abcam), β-actin (ab8227; 1:500; Abcam). .. Subsequently, the membranes were incubated with secondary goat monoclonal (RMG01) to rabbit IgG Fab region (Biotinylated; ab222772; 1:10,000; Abcam) at room temperature for 2 h, and proteins were detected using enhanced chemiluminescence (Pierce™ ECL Western Blotting Substrate; Thermo Fisher Scientific, Inc.).

    SDS Page:

    Article Title: miR-204 functions as a tumor suppressor gene, at least partly by suppressing CYP27A1 in glioblastoma
    Article Snippet: .. Following heating at 100°C for 10 min in the presence of a loading buffer, equal amounts of protein lysates (50 µg) were separated using 10% SDS-PAGE (Bio-West Inc., Logan, UT, USA) at 100 V for 1 h, and transferred onto Invitrogen nitrocellulose membranes (Thermo Fisher Scientific, Inc.) at 120 V for 1 h. Following blocking with 5% skimmed milk (diluted with PBS), the membranes were incubated overnight at 4°C with the following primary antibodies: CYP27A1 (ab126785; 1:500; Abcam, Cambridge, MA, USA), c-myc (ab32072; 1:500; Abcam), RB (ab181616; 1:500; Abcam), Ki-67 (ab15580; 1:500; Abcam), CDK2 (ab32147; 1:500; Abcam), p21 (ab109520; 1:500; Abcam), p53 (ab32049; 1:500; Abcam), PDCD4 (ab51495; 1:500; Abcam), SOX2 (ab92494; 1:500; Abcam), β-actin (ab8227; 1:500; Abcam). .. Subsequently, the membranes were incubated with secondary goat monoclonal (RMG01) to rabbit IgG Fab region (Biotinylated; ab222772; 1:10,000; Abcam) at room temperature for 2 h, and proteins were detected using enhanced chemiluminescence (Pierce™ ECL Western Blotting Substrate; Thermo Fisher Scientific, Inc.).

    Concentration Assay:

    Article Title: Transcriptionally dynamic progenitor populations organised around a stable niche drive axial patterning
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    Incubation:

    Article Title: MiR-208a stimulates the cocktail of SOX2 and β-catenin to inhibit the let-7 induction of self-renewal repression of breast cancer stem cells and formed miR208a/let-7 feedback loop via LIN28 and DICER1
    Article Snippet: .. Immunofluorescence staining used SOX2 (1:200, ab92494, Abcam, USA) and LIN28 (1:100, AP1485A, ABGENT, USA); the slide was incubated with a secondary antibody for 30 min with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 h at room temperature. .. Cells were counterstained with DRAQ5 (1:10000; 4084; Cell Signaling, China) to show nuclei.

    Article Title: miR-204 functions as a tumor suppressor gene, at least partly by suppressing CYP27A1 in glioblastoma
    Article Snippet: .. Following heating at 100°C for 10 min in the presence of a loading buffer, equal amounts of protein lysates (50 µg) were separated using 10% SDS-PAGE (Bio-West Inc., Logan, UT, USA) at 100 V for 1 h, and transferred onto Invitrogen nitrocellulose membranes (Thermo Fisher Scientific, Inc.) at 120 V for 1 h. Following blocking with 5% skimmed milk (diluted with PBS), the membranes were incubated overnight at 4°C with the following primary antibodies: CYP27A1 (ab126785; 1:500; Abcam, Cambridge, MA, USA), c-myc (ab32072; 1:500; Abcam), RB (ab181616; 1:500; Abcam), Ki-67 (ab15580; 1:500; Abcam), CDK2 (ab32147; 1:500; Abcam), p21 (ab109520; 1:500; Abcam), p53 (ab32049; 1:500; Abcam), PDCD4 (ab51495; 1:500; Abcam), SOX2 (ab92494; 1:500; Abcam), β-actin (ab8227; 1:500; Abcam). .. Subsequently, the membranes were incubated with secondary goat monoclonal (RMG01) to rabbit IgG Fab region (Biotinylated; ab222772; 1:10,000; Abcam) at room temperature for 2 h, and proteins were detected using enhanced chemiluminescence (Pierce™ ECL Western Blotting Substrate; Thermo Fisher Scientific, Inc.).

    Staining:

    Article Title: MiR-208a stimulates the cocktail of SOX2 and β-catenin to inhibit the let-7 induction of self-renewal repression of breast cancer stem cells and formed miR208a/let-7 feedback loop via LIN28 and DICER1
    Article Snippet: .. Immunofluorescence staining used SOX2 (1:200, ab92494, Abcam, USA) and LIN28 (1:100, AP1485A, ABGENT, USA); the slide was incubated with a secondary antibody for 30 min with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 h at room temperature. .. Cells were counterstained with DRAQ5 (1:10000; 4084; Cell Signaling, China) to show nuclei.

    Immunofluorescence:

    Article Title: MiR-208a stimulates the cocktail of SOX2 and β-catenin to inhibit the let-7 induction of self-renewal repression of breast cancer stem cells and formed miR208a/let-7 feedback loop via LIN28 and DICER1
    Article Snippet: .. Immunofluorescence staining used SOX2 (1:200, ab92494, Abcam, USA) and LIN28 (1:100, AP1485A, ABGENT, USA); the slide was incubated with a secondary antibody for 30 min with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 h at room temperature. .. Cells were counterstained with DRAQ5 (1:10000; 4084; Cell Signaling, China) to show nuclei.

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  • sox2  (Abcam)
    96
    Abcam sox2
    miR-21 suppresses <t>Sox2</t> expression in E14Tg2a.4 mESCs
    Sox2, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sox2/product/Abcam
    Average 96 stars, based on 133 article reviews
    Price from $9.99 to $1999.99
    sox2 - by Bioz Stars, 2020-07
    96/100 stars
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    86
    Abcam monoclonal mouse anti human sox2
    Western blot analysis of <t>SOX2,</t> NANOG, and OCT protein. (A) Relative levels of SOX2, NANOG, and OCT4 protein in normal endometrium (n = 6) and paired eutopic and ectopic endometrium (n = 13). Double-asterisk values are significantly different from single-asterisk values. Asterisks denote significant differences between eutopic or ectopic endometrium and normal endometrium. Connecting lines denote significant differences between eutopic and ectopic endometrium. P
    Monoclonal Mouse Anti Human Sox2, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal mouse anti human sox2/product/Abcam
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    Price from $9.99 to $1999.99
    monoclonal mouse anti human sox2 - by Bioz Stars, 2020-07
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    Image Search Results


    miR-21 suppresses Sox2 expression in E14Tg2a.4 mESCs

    Journal: Stem cell research

    Article Title: REST-miR-21-SOX2 axis maintains pluripotency in E14Tg2a.4 embryonic stem cells

    doi: 10.1016/j.scr.2015.05.003

    Figure Lengend Snippet: miR-21 suppresses Sox2 expression in E14Tg2a.4 mESCs

    Article Snippet: Whole-cell extracts were prepared, and approximately 10-30 μg of protein was resolved on SDS-PAGE using antibodies against REST (#07-579, Upstate), Oct4 (#ab19857, Abcam), anti-hemaglutinin (HA) (H9658, Sigma), Sox2 (#ab15830, Abcam; #2748, Cell Signaling), β-actin (A5316, Sigma), and α-tubulin (#MMS-407R, Covance). mESCs were transfected with pre-miR21 (Pre-miR miRNA precursor molecule; AM17100, Ambion) or control, as described earlier ( ). mESCs were transfected using AMAXA nucleofection reagent.

    Techniques: Expressing

    Exogenous Sox2 with deleted miR-21 binding site counters the suppression of self-renewal by miR-21 in mESCs

    Journal: Stem cell research

    Article Title: REST-miR-21-SOX2 axis maintains pluripotency in E14Tg2a.4 embryonic stem cells

    doi: 10.1016/j.scr.2015.05.003

    Figure Lengend Snippet: Exogenous Sox2 with deleted miR-21 binding site counters the suppression of self-renewal by miR-21 in mESCs

    Article Snippet: Whole-cell extracts were prepared, and approximately 10-30 μg of protein was resolved on SDS-PAGE using antibodies against REST (#07-579, Upstate), Oct4 (#ab19857, Abcam), anti-hemaglutinin (HA) (H9658, Sigma), Sox2 (#ab15830, Abcam; #2748, Cell Signaling), β-actin (A5316, Sigma), and α-tubulin (#MMS-407R, Covance). mESCs were transfected with pre-miR21 (Pre-miR miRNA precursor molecule; AM17100, Ambion) or control, as described earlier ( ). mESCs were transfected using AMAXA nucleofection reagent.

    Techniques: Binding Assay

    Effect of LGR5 on intracranial tumor growth and overall survival time of xenograft mice. a T2W images of intracranial tumor in coronal views scanned using 7.0 T MRI on days 12, 25 and 39 after U251-GSCs injection. b The intracranial tumor growth curve of the shLGR5 ( n = 5), Wnt-C59 ( n = 5) and shCtrl ( n = 5) U251-GSCs. Error bars represent the mean ± SD ( n = 5, two-way ANOVA). c Overall survival time (OS) of xenograft mice in the shLGR5 ( n = 5), Wnt-C59 ( n = 5) and shCtrl ( n = 5) U251-GSCs analyzed by Log-rank test. d IHC staining and quantitative analyses of LGR5, Ki67, Active-β-catenin, SOX2 and CD44 (magnification × 200). e Spearman correlation analysis of LGR5 and Active-β-catenin between shLGR5 and shCtrl U251-GSCs ( P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: LGR5, a novel functional glioma stem cell marker, promotes EMT by activating the Wnt/β-catenin pathway and predicts poor survival of glioma patients

    doi: 10.1186/s13046-018-0864-6

    Figure Lengend Snippet: Effect of LGR5 on intracranial tumor growth and overall survival time of xenograft mice. a T2W images of intracranial tumor in coronal views scanned using 7.0 T MRI on days 12, 25 and 39 after U251-GSCs injection. b The intracranial tumor growth curve of the shLGR5 ( n = 5), Wnt-C59 ( n = 5) and shCtrl ( n = 5) U251-GSCs. Error bars represent the mean ± SD ( n = 5, two-way ANOVA). c Overall survival time (OS) of xenograft mice in the shLGR5 ( n = 5), Wnt-C59 ( n = 5) and shCtrl ( n = 5) U251-GSCs analyzed by Log-rank test. d IHC staining and quantitative analyses of LGR5, Ki67, Active-β-catenin, SOX2 and CD44 (magnification × 200). e Spearman correlation analysis of LGR5 and Active-β-catenin between shLGR5 and shCtrl U251-GSCs ( P

    Article Snippet: All antibodies used were as follows: LGR5 (#TA502948; OriGene), Ki67 (#ab16667; Abcam), CD44 (#15675–1-AP, Proteintech), SOX2 (#ab92494; Abcam), Non-phospho (Active) β-catenin Ser33/37/Thr41 (#8814; Cell Signaling) and N-cadherin (#bs-1172R; Bioss).

    Techniques: Mouse Assay, Magnetic Resonance Imaging, Injection, Immunohistochemistry, Staining

    Relationship between LGR5 and reported CSCs markers and stem cell genes. a Analyses of CD133, CD44, CD90, CD24, and EpCAM expression in LGR5 + and LGR5 − U251 glioma cells and in LGR5 + and LGR5 − 8591 human primary glioma cells by FCM. b U251-GSCs and 8591-GSCs from U251 and 8591 human primary glioma cells through serum-free enrichment (top, scale bar = 100 μm) and immunofluorescence double staining for LGR5 and SOX2 (bottom, scale bar = 50 μm). c The expression levels of LGR5, CSCs markers (CD133, CD44, CD24 and EpCAM) and stem cell genes (SOX2, OCT4 and Nanog) in shLGR5 and shCtrl GSCs determined by Western blot (top). The protein intensity of these markers were determined by densitometric analysis and normalized to the relevant β-actin value. Error bars represent the mean ± SD of three independent experiments (bottom). *, P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: LGR5, a novel functional glioma stem cell marker, promotes EMT by activating the Wnt/β-catenin pathway and predicts poor survival of glioma patients

    doi: 10.1186/s13046-018-0864-6

    Figure Lengend Snippet: Relationship between LGR5 and reported CSCs markers and stem cell genes. a Analyses of CD133, CD44, CD90, CD24, and EpCAM expression in LGR5 + and LGR5 − U251 glioma cells and in LGR5 + and LGR5 − 8591 human primary glioma cells by FCM. b U251-GSCs and 8591-GSCs from U251 and 8591 human primary glioma cells through serum-free enrichment (top, scale bar = 100 μm) and immunofluorescence double staining for LGR5 and SOX2 (bottom, scale bar = 50 μm). c The expression levels of LGR5, CSCs markers (CD133, CD44, CD24 and EpCAM) and stem cell genes (SOX2, OCT4 and Nanog) in shLGR5 and shCtrl GSCs determined by Western blot (top). The protein intensity of these markers were determined by densitometric analysis and normalized to the relevant β-actin value. Error bars represent the mean ± SD of three independent experiments (bottom). *, P

    Article Snippet: All antibodies used were as follows: LGR5 (#TA502948; OriGene), Ki67 (#ab16667; Abcam), CD44 (#15675–1-AP, Proteintech), SOX2 (#ab92494; Abcam), Non-phospho (Active) β-catenin Ser33/37/Thr41 (#8814; Cell Signaling) and N-cadherin (#bs-1172R; Bioss).

    Techniques: Expressing, Immunofluorescence, Double Staining, Western Blot

    The positive correlation between SOX2/β-catenin and LIN28 and their clinical value in evaluating prognosis of patients with breast cancer Representative examples of immunohistochemical staining for LIN-28 A. SOX2 B. and β-catenin C. in each of the clinical stages of breast cancers as indicated. Stage I had the lowest expression of LIN28, SOX2, and β-catenin. D. Quantification of LIN-28, SOX2, and β-catenin relative immunostaining intensity for each clinical stage of breast cancers. Data is shown as mean ± SEM. E. Relative expression levels of SOX2 and LIN28, β-catenin and LIN28, and SOX2 and β-catenin, with correlation coefficients shown in figure.

    Journal: Oncotarget

    Article Title: MiR-208a stimulates the cocktail of SOX2 and β-catenin to inhibit the let-7 induction of self-renewal repression of breast cancer stem cells and formed miR208a/let-7 feedback loop via LIN28 and DICER1

    doi:

    Figure Lengend Snippet: The positive correlation between SOX2/β-catenin and LIN28 and their clinical value in evaluating prognosis of patients with breast cancer Representative examples of immunohistochemical staining for LIN-28 A. SOX2 B. and β-catenin C. in each of the clinical stages of breast cancers as indicated. Stage I had the lowest expression of LIN28, SOX2, and β-catenin. D. Quantification of LIN-28, SOX2, and β-catenin relative immunostaining intensity for each clinical stage of breast cancers. Data is shown as mean ± SEM. E. Relative expression levels of SOX2 and LIN28, β-catenin and LIN28, and SOX2 and β-catenin, with correlation coefficients shown in figure.

    Article Snippet: Immunofluorescence staining used SOX2 (1:200, ab92494, Abcam, USA) and LIN28 (1:100, AP1485A, ABGENT, USA); the slide was incubated with a secondary antibody for 30 min with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 h at room temperature.

    Techniques: Immunohistochemistry, Staining, Expressing, Immunostaining

    MiR-208a promotes LIN28 via regulations of SOX2 and β-catenin A. SOX2, β-catenin and LIN28 are upregulated in mammospheres from ZR75–1 and MM-231 cells, compared to adherent cells. B. miR-208a stimulates LIN28 expression via SOX2 and β-catenin partially, and the inhibition of SOX2 or β-catenin could only partially functions of miR-208a in induction of LIN28. C. siRNAs of SOX2 and β-catenin together effectively inhibited LIN28 expression in miR-208a overexpressed ZR75–1 and MM-231 cells. D. SOX2 and LIN28 increased in miR-208a overexpressed breast cancer stem cells. E. The inhibition of SOX2 or β-catenin decreased the ratio of ALDH1+ stem cells; this inhibition was significantly greater when both SOX2 and β-catenin siRNA was used, reducing the ratio of ALDH1+ cells to control levels. F. No significant differences of ratios of ALDH1+ cells were detected in groups of LIN28 inhibition and SOX2/β-catenin inhibition.

    Journal: Oncotarget

    Article Title: MiR-208a stimulates the cocktail of SOX2 and β-catenin to inhibit the let-7 induction of self-renewal repression of breast cancer stem cells and formed miR208a/let-7 feedback loop via LIN28 and DICER1

    doi:

    Figure Lengend Snippet: MiR-208a promotes LIN28 via regulations of SOX2 and β-catenin A. SOX2, β-catenin and LIN28 are upregulated in mammospheres from ZR75–1 and MM-231 cells, compared to adherent cells. B. miR-208a stimulates LIN28 expression via SOX2 and β-catenin partially, and the inhibition of SOX2 or β-catenin could only partially functions of miR-208a in induction of LIN28. C. siRNAs of SOX2 and β-catenin together effectively inhibited LIN28 expression in miR-208a overexpressed ZR75–1 and MM-231 cells. D. SOX2 and LIN28 increased in miR-208a overexpressed breast cancer stem cells. E. The inhibition of SOX2 or β-catenin decreased the ratio of ALDH1+ stem cells; this inhibition was significantly greater when both SOX2 and β-catenin siRNA was used, reducing the ratio of ALDH1+ cells to control levels. F. No significant differences of ratios of ALDH1+ cells were detected in groups of LIN28 inhibition and SOX2/β-catenin inhibition.

    Article Snippet: Immunofluorescence staining used SOX2 (1:200, ab92494, Abcam, USA) and LIN28 (1:100, AP1485A, ABGENT, USA); the slide was incubated with a secondary antibody for 30 min with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1 h at room temperature.

    Techniques: Expressing, Inhibition

    Western blot analysis of SOX2, NANOG, and OCT protein. (A) Relative levels of SOX2, NANOG, and OCT4 protein in normal endometrium (n = 6) and paired eutopic and ectopic endometrium (n = 13). Double-asterisk values are significantly different from single-asterisk values. Asterisks denote significant differences between eutopic or ectopic endometrium and normal endometrium. Connecting lines denote significant differences between eutopic and ectopic endometrium. P

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Increased expression of the pluripotency markers sex-determining region Y-box 2 and Nanog homeobox in ovarian endometriosis

    doi: 10.1186/1477-7827-12-42

    Figure Lengend Snippet: Western blot analysis of SOX2, NANOG, and OCT protein. (A) Relative levels of SOX2, NANOG, and OCT4 protein in normal endometrium (n = 6) and paired eutopic and ectopic endometrium (n = 13). Double-asterisk values are significantly different from single-asterisk values. Asterisks denote significant differences between eutopic or ectopic endometrium and normal endometrium. Connecting lines denote significant differences between eutopic and ectopic endometrium. P

    Article Snippet: Immunohistochemistry Immunohistochemical staining was performed using monoclonal mouse anti-human SOX2 (ab75485, Abcam, Cambridge, MA, USA), polyclonal rabbit anti-human NANOG (ab80892, Abcam), and rabbit anti-human OCT4 (ab18976, Abcam) as primary antibodies.

    Techniques: Western Blot

    Quantitative real-time PCR analysis of SOX2 , NANOG , and OCT4 mRNA. Analysis was carried out on normal endometrium (n = 10) and paired eutopic and ectopic endometrium specimens (n = 13). Double-asterisk values are significantly different from single-asterisk values. Asterisks denote significant differences between eutopic or ectopic endometrium and normal endometrium. Connecting lines denote significant differences between eutopic and ectopic endometrium. P

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Increased expression of the pluripotency markers sex-determining region Y-box 2 and Nanog homeobox in ovarian endometriosis

    doi: 10.1186/1477-7827-12-42

    Figure Lengend Snippet: Quantitative real-time PCR analysis of SOX2 , NANOG , and OCT4 mRNA. Analysis was carried out on normal endometrium (n = 10) and paired eutopic and ectopic endometrium specimens (n = 13). Double-asterisk values are significantly different from single-asterisk values. Asterisks denote significant differences between eutopic or ectopic endometrium and normal endometrium. Connecting lines denote significant differences between eutopic and ectopic endometrium. P

    Article Snippet: Immunohistochemistry Immunohistochemical staining was performed using monoclonal mouse anti-human SOX2 (ab75485, Abcam, Cambridge, MA, USA), polyclonal rabbit anti-human NANOG (ab80892, Abcam), and rabbit anti-human OCT4 (ab18976, Abcam) as primary antibodies.

    Techniques: Real-time Polymerase Chain Reaction

    SOX2, NANOG, and OCT4 immunohistochemical staining. Paraffin-embedded normal (n = 6) and paired eutopic and ectopic endometrium specimens (n = 7) were examined. SOX2, NANOG, and OCT4 immunostaining in normal endometrium (C, H, M) of control and in eutopic endometrium (D, I, N) and ectopic endometrium (E, J, O) of endometriosis, and positive control (A, F, K) and negative control (B, G, L) were revealed, respectively (×400 magnification). Arrows denote positive staining.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Increased expression of the pluripotency markers sex-determining region Y-box 2 and Nanog homeobox in ovarian endometriosis

    doi: 10.1186/1477-7827-12-42

    Figure Lengend Snippet: SOX2, NANOG, and OCT4 immunohistochemical staining. Paraffin-embedded normal (n = 6) and paired eutopic and ectopic endometrium specimens (n = 7) were examined. SOX2, NANOG, and OCT4 immunostaining in normal endometrium (C, H, M) of control and in eutopic endometrium (D, I, N) and ectopic endometrium (E, J, O) of endometriosis, and positive control (A, F, K) and negative control (B, G, L) were revealed, respectively (×400 magnification). Arrows denote positive staining.

    Article Snippet: Immunohistochemistry Immunohistochemical staining was performed using monoclonal mouse anti-human SOX2 (ab75485, Abcam, Cambridge, MA, USA), polyclonal rabbit anti-human NANOG (ab80892, Abcam), and rabbit anti-human OCT4 (ab18976, Abcam) as primary antibodies.

    Techniques: Immunohistochemistry, Staining, Immunostaining, Positive Control, Negative Control