Structured Review

Becton Dickinson anti sox2
Proliferation in the embryonic cochleae of Lgr4 deficient mice. (A–B′′) In the cochleae from heterozygous and homozygous Lgr4-LacZ mice, CCND1 (green) expression increased in the mid-basal turn, where CCND1 merged with red signal of <t>SOX2</t> (arrows) in the pro-sensory domain. (C–D′′) The proliferation marker, EdU (red), was incorporated into pro-sensory cells (arrows) positive for SOX2 (green) in the mid-basal turn of the cochlea from Lgr4 deficient mice. Cell nuclei were counterstained with DAPI (blue), scale bars indicate 20 μm.
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1) Product Images from "LGR4 and LGR5 Regulate Hair Cell Differentiation in the Sensory Epithelium of the Developing Mouse Cochlea"

Article Title: LGR4 and LGR5 Regulate Hair Cell Differentiation in the Sensory Epithelium of the Developing Mouse Cochlea

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2016.00186

Proliferation in the embryonic cochleae of Lgr4 deficient mice. (A–B′′) In the cochleae from heterozygous and homozygous Lgr4-LacZ mice, CCND1 (green) expression increased in the mid-basal turn, where CCND1 merged with red signal of SOX2 (arrows) in the pro-sensory domain. (C–D′′) The proliferation marker, EdU (red), was incorporated into pro-sensory cells (arrows) positive for SOX2 (green) in the mid-basal turn of the cochlea from Lgr4 deficient mice. Cell nuclei were counterstained with DAPI (blue), scale bars indicate 20 μm.
Figure Legend Snippet: Proliferation in the embryonic cochleae of Lgr4 deficient mice. (A–B′′) In the cochleae from heterozygous and homozygous Lgr4-LacZ mice, CCND1 (green) expression increased in the mid-basal turn, where CCND1 merged with red signal of SOX2 (arrows) in the pro-sensory domain. (C–D′′) The proliferation marker, EdU (red), was incorporated into pro-sensory cells (arrows) positive for SOX2 (green) in the mid-basal turn of the cochlea from Lgr4 deficient mice. Cell nuclei were counterstained with DAPI (blue), scale bars indicate 20 μm.

Techniques Used: Mouse Assay, Expressing, Marker

The distribution of LGR4 in the developing mouse cochlea. (A) At E14.5, strong LGR4 staining was detected in the cochlear duct and spiral ganglion cells (white arrowhead). LGR4 was detected in the pro-sensory domain, where it overlapped with SOX2 (white arrow), and non-sensory domains lateral and medial to the pro-sensory domain. (B) At E17, LGR4 (green) was detected in IHCs and OHCs (asterisks) and their surrounding supporting cells. Hair cells were labeled with phalloidin (F-actin) (red). (C) At P1, LGR4 expression was observed in the sensory epithelium (arrowhead) and spiral ganglion cells (arrow), from which projected TUJ1-positive nerves (red). (D) At P7, LGR4 was observed in hair cells (asterisks) that co-expressed Prestin (PRES) and in the surrounding supporting cells: Deiters’ cells (arrowheads) and IPCs and OPCs. (E–G) At P21, LGR4 was down-regulated in the OHCs and pillar cells, but was still expressed in the IHCs (arrowhead), Deiters’ cells (arrows), which were also positive for acetylated tubulin (ACTBA), and phalangeal processes of Deiters’ cells (arrowheads). LGR4 signals were also observed in the spiral ganglion cells. (H) At P42, LGR4 was detected in Deiters’ cells and their phalangeal processes. Cell nuclei were counterstained with DAPI (blue), asterisks indicate hair cells, scale bars indicate 20 μm in the picture (B,D,E,G,H) and 50 μm in (A,C,F) . (I,J) Cochlear cryosections from Lgr4-LacZ mice at the age of P21 were stained for β-galactosidase activity. Expression was seen in Deiters’ cells (black arrow) and their phalangeal processes (gray arrow), IHCs (black arrowhead), and spiral ganglion cells.
Figure Legend Snippet: The distribution of LGR4 in the developing mouse cochlea. (A) At E14.5, strong LGR4 staining was detected in the cochlear duct and spiral ganglion cells (white arrowhead). LGR4 was detected in the pro-sensory domain, where it overlapped with SOX2 (white arrow), and non-sensory domains lateral and medial to the pro-sensory domain. (B) At E17, LGR4 (green) was detected in IHCs and OHCs (asterisks) and their surrounding supporting cells. Hair cells were labeled with phalloidin (F-actin) (red). (C) At P1, LGR4 expression was observed in the sensory epithelium (arrowhead) and spiral ganglion cells (arrow), from which projected TUJ1-positive nerves (red). (D) At P7, LGR4 was observed in hair cells (asterisks) that co-expressed Prestin (PRES) and in the surrounding supporting cells: Deiters’ cells (arrowheads) and IPCs and OPCs. (E–G) At P21, LGR4 was down-regulated in the OHCs and pillar cells, but was still expressed in the IHCs (arrowhead), Deiters’ cells (arrows), which were also positive for acetylated tubulin (ACTBA), and phalangeal processes of Deiters’ cells (arrowheads). LGR4 signals were also observed in the spiral ganglion cells. (H) At P42, LGR4 was detected in Deiters’ cells and their phalangeal processes. Cell nuclei were counterstained with DAPI (blue), asterisks indicate hair cells, scale bars indicate 20 μm in the picture (B,D,E,G,H) and 50 μm in (A,C,F) . (I,J) Cochlear cryosections from Lgr4-LacZ mice at the age of P21 were stained for β-galactosidase activity. Expression was seen in Deiters’ cells (black arrow) and their phalangeal processes (gray arrow), IHCs (black arrowhead), and spiral ganglion cells.

Techniques Used: Staining, Labeling, Expressing, Mouse Assay, Activity Assay

Proliferation in the cochleae of E14.5 Lgr5-eGFP embryos. (A–B′′) SOX2-positive pro-sensory cells (green, arrow) did not incorporate EdU (red). Cell nuclei were counterstained with DAPI (blue), scale bars indicate 25 μm.
Figure Legend Snippet: Proliferation in the cochleae of E14.5 Lgr5-eGFP embryos. (A–B′′) SOX2-positive pro-sensory cells (green, arrow) did not incorporate EdU (red). Cell nuclei were counterstained with DAPI (blue), scale bars indicate 25 μm.

Techniques Used:

2) Product Images from "Fixed single-cell transcriptomic characterization of human radial glial diversity"

Article Title: Fixed single-cell transcriptomic characterization of human radial glial diversity

Journal: Nature methods

doi: 10.1038/nmeth.3629

FRISCR allows profiling of primary human cortical progenitors. Color scheme throughout figure is shown in b . ( a ) FACS plots show gating strategy and frequency of gated cells compared to total cortical cells from three independent experiments. Left plot includes only G 0 -G 1 singlet cortical cells (gated with DAPI) and right plot shows only the SOX2 + PAX6 + cells gated on the left plot. ( b ) Barplots show the frequency of indicated genes detected per cell. Only cells with > 10,000 mapped reads, > 20% mRNA mapping, and GAPDH expression ( n = 157 SP and n = 29 SPE cells) were included in this analysis.
Figure Legend Snippet: FRISCR allows profiling of primary human cortical progenitors. Color scheme throughout figure is shown in b . ( a ) FACS plots show gating strategy and frequency of gated cells compared to total cortical cells from three independent experiments. Left plot includes only G 0 -G 1 singlet cortical cells (gated with DAPI) and right plot shows only the SOX2 + PAX6 + cells gated on the left plot. ( b ) Barplots show the frequency of indicated genes detected per cell. Only cells with > 10,000 mapped reads, > 20% mRNA mapping, and GAPDH expression ( n = 157 SP and n = 29 SPE cells) were included in this analysis.

Techniques Used: FACS, Expressing

Confirmation of vRG and oRG markers. ( a ) Grayscale ( left ) or merged ( right ) micrographs of germinal zone showing co-staining of SOX2 (magenta) and ANXA1, CRYAB or HOPX (green). Tissue is 16 PCW cortex, scales are 100 µm. Abbreviations: marginal zone (MZ), cortical plate (CP), subplate/intermediate zone (SP/IZ), outer subventricular zone (oSZ), inner subventricular zone (iSZ), and ventricular zone (VZ). ( b ) HOPX expression in the VZ; scale is 50 µm. Co-expression with SOX2 (arrowhead), SOX2-only cells (arrow), and HOPX + apical endfoot are indicated (asterisk). ( c ) A mitotic cell co-stained with SOX2, HOPX and phospho-VIM shows basal cell process (asterisk) and cell body (arrowhead); scale is 20 µm. ( d ) Full cortical thickness (left), and insets (right) from germinal zone showing co-staining of SOX2 (magenta) and HOPX (green). Scales are 100 µm. ( e ) Quantitation of SOX2 + HOPX + or SOX2 + HOPX − cells in oSZ that are Ki67 + . ( f ) Quantification of SOX2 + ANXA1 + or SOX2 + ANXA1 − cells in VZ that are Ki67 + . Data is mean ± SD for ( e ) five or ( f ) six human tissues at 14–19 PCW; ** P
Figure Legend Snippet: Confirmation of vRG and oRG markers. ( a ) Grayscale ( left ) or merged ( right ) micrographs of germinal zone showing co-staining of SOX2 (magenta) and ANXA1, CRYAB or HOPX (green). Tissue is 16 PCW cortex, scales are 100 µm. Abbreviations: marginal zone (MZ), cortical plate (CP), subplate/intermediate zone (SP/IZ), outer subventricular zone (oSZ), inner subventricular zone (iSZ), and ventricular zone (VZ). ( b ) HOPX expression in the VZ; scale is 50 µm. Co-expression with SOX2 (arrowhead), SOX2-only cells (arrow), and HOPX + apical endfoot are indicated (asterisk). ( c ) A mitotic cell co-stained with SOX2, HOPX and phospho-VIM shows basal cell process (asterisk) and cell body (arrowhead); scale is 20 µm. ( d ) Full cortical thickness (left), and insets (right) from germinal zone showing co-staining of SOX2 (magenta) and HOPX (green). Scales are 100 µm. ( e ) Quantitation of SOX2 + HOPX + or SOX2 + HOPX − cells in oSZ that are Ki67 + . ( f ) Quantification of SOX2 + ANXA1 + or SOX2 + ANXA1 − cells in VZ that are Ki67 + . Data is mean ± SD for ( e ) five or ( f ) six human tissues at 14–19 PCW; ** P

Techniques Used: Staining, Expressing, Quantitation Assay

Human cortical progenitors are diverse and intermixed during development. ( a ) Model of the progenitor compartment shows a mixture of ventricular radial glial cells (vRG-light blue), outer RGs (oRGs-purple), intermediate progenitors (IPCs-orange) and other mixed cortical cell types (gray). Known markers for each cell type are shown below. Note RGs identified by antibody staining are called SP (SOX + PAX + EOMES − -dark blue nuclei), and IPCs are called SPE (SOX + PAX + EOMES + -dark orange nuclei). ( b ) Immunocytochemistry images of 19 PCW germinal zones. Top left : Low magnification images stitched together ( left ), or individual micrograph ( right ) of the VZ, iSZ, and oSZ stained by DAPI, EOMES, PAX6, and SOX2 with the colors indicated. Scale bars are 100 µm. High magnification micrograph of oSZ region ( middle ) and VZ/iSZ region ( bottom ) show SP (arrow with filled arrowhead) and SPE (arrowhead) cells. Note many cells the VZ, iSZ, and oSZ lack progenitor markers and are unknown cell types (arrows with open arrowhead). Scale bars are 50 µm. Tables describing human germinal zone regions and symbols ( c ), and common human cortical progenitors, their germinal zone locations, and their flow cytometry (FACS) phenotypes ( d ).
Figure Legend Snippet: Human cortical progenitors are diverse and intermixed during development. ( a ) Model of the progenitor compartment shows a mixture of ventricular radial glial cells (vRG-light blue), outer RGs (oRGs-purple), intermediate progenitors (IPCs-orange) and other mixed cortical cell types (gray). Known markers for each cell type are shown below. Note RGs identified by antibody staining are called SP (SOX + PAX + EOMES − -dark blue nuclei), and IPCs are called SPE (SOX + PAX + EOMES + -dark orange nuclei). ( b ) Immunocytochemistry images of 19 PCW germinal zones. Top left : Low magnification images stitched together ( left ), or individual micrograph ( right ) of the VZ, iSZ, and oSZ stained by DAPI, EOMES, PAX6, and SOX2 with the colors indicated. Scale bars are 100 µm. High magnification micrograph of oSZ region ( middle ) and VZ/iSZ region ( bottom ) show SP (arrow with filled arrowhead) and SPE (arrowhead) cells. Note many cells the VZ, iSZ, and oSZ lack progenitor markers and are unknown cell types (arrows with open arrowhead). Scale bars are 50 µm. Tables describing human germinal zone regions and symbols ( c ), and common human cortical progenitors, their germinal zone locations, and their flow cytometry (FACS) phenotypes ( d ).

Techniques Used: Staining, Immunocytochemistry, Flow Cytometry, Cytometry, FACS

3) Product Images from "Lovastatin Decreases the Expression of CD133 and Influences the Differentiation Potential of Human Embryonic Stem Cells"

Article Title: Lovastatin Decreases the Expression of CD133 and Influences the Differentiation Potential of Human Embryonic Stem Cells

Journal: Stem Cells International

doi: 10.1155/2016/1580701

Expression of differentiation markers in hES cells during differentiation into ectodermal, mesodermal, or endodermal lineages after treatment with lovastatin. (a) Expression of the ectodermal lineage markers SOX1 and OTX2 in comparison to the expression of OCT4 and SOX2. (b) Expression of the endodermal lineage markers GATA4, FOXA2, and SOX17 in comparison to OCT4 expression. (c) Expression of the mesodermal lineage markers brachyury and HAND1 and in comparison to the expression of OCT4. (d) The mean number of cells ± SD ( n = 5) expressing ectodermal (EC), mesodermal (ME), or endodermal (EN) lineage markers.
Figure Legend Snippet: Expression of differentiation markers in hES cells during differentiation into ectodermal, mesodermal, or endodermal lineages after treatment with lovastatin. (a) Expression of the ectodermal lineage markers SOX1 and OTX2 in comparison to the expression of OCT4 and SOX2. (b) Expression of the endodermal lineage markers GATA4, FOXA2, and SOX17 in comparison to OCT4 expression. (c) Expression of the mesodermal lineage markers brachyury and HAND1 and in comparison to the expression of OCT4. (d) The mean number of cells ± SD ( n = 5) expressing ectodermal (EC), mesodermal (ME), or endodermal (EN) lineage markers.

Techniques Used: Expressing

CD133 expression was crucial for proper differentiation into embryoid bodies. CD133+ and CD133− cell subpopulations were enriched using the magnetic beads separation method from untreated hES cells and the formation of EB was initiated. (a) Morphology of the formed structures on day five. (b) Formed EB and cell structures were analysed for the expression of CD184, nestin, and SOX2.
Figure Legend Snippet: CD133 expression was crucial for proper differentiation into embryoid bodies. CD133+ and CD133− cell subpopulations were enriched using the magnetic beads separation method from untreated hES cells and the formation of EB was initiated. (a) Morphology of the formed structures on day five. (b) Formed EB and cell structures were analysed for the expression of CD184, nestin, and SOX2.

Techniques Used: Expressing, Magnetic Beads

Characterization of untreated and lovastatin treated EB cells for ectodermal differentiation markers. (a) Single cell suspensions from EB were analysed for the expression of CD184 (CXCR4), nestin, SOX2, and OTX2. (b) The mean number of cells ± SD ( n = 4) expressing the markers.
Figure Legend Snippet: Characterization of untreated and lovastatin treated EB cells for ectodermal differentiation markers. (a) Single cell suspensions from EB were analysed for the expression of CD184 (CXCR4), nestin, SOX2, and OTX2. (b) The mean number of cells ± SD ( n = 4) expressing the markers.

Techniques Used: Expressing

Lovastatin treatment differently affects the expression of pluripotency markers in hES cells. (a) Flow cytometric analysis of the expression of the pluripotency markers NANOG, OCT4, and SOX2 in hES cells treated with lovastatin (20 μ M lovastatin for 48 h) and those untreated. Fixed and permeabilised cells were stained with anti-SSEA-3 (Alexa Fluor 488 conjugate), anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) The morphological changes in colony structure of lovastatin treated hES cells. (c) The number of cells expressing pluripotency markers. Results are shown as mean ± SD ( n = 5). (d) The number of cells in one well after 48 h of treatment with lovastatin. (e) Changes in SSEA-3 expression in untreated and lovastatin treated (10–40 μ M for 24 h) hES cells.
Figure Legend Snippet: Lovastatin treatment differently affects the expression of pluripotency markers in hES cells. (a) Flow cytometric analysis of the expression of the pluripotency markers NANOG, OCT4, and SOX2 in hES cells treated with lovastatin (20 μ M lovastatin for 48 h) and those untreated. Fixed and permeabilised cells were stained with anti-SSEA-3 (Alexa Fluor 488 conjugate), anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) The morphological changes in colony structure of lovastatin treated hES cells. (c) The number of cells expressing pluripotency markers. Results are shown as mean ± SD ( n = 5). (d) The number of cells in one well after 48 h of treatment with lovastatin. (e) Changes in SSEA-3 expression in untreated and lovastatin treated (10–40 μ M for 24 h) hES cells.

Techniques Used: Expressing, Flow Cytometry, Staining

4) Product Images from "Functional development of mechanosensitive hair cells in stem cell-derived organoids parallels native vestibular hair cells"

Article Title: Functional development of mechanosensitive hair cells in stem cell-derived organoids parallels native vestibular hair cells

Journal: Nature Communications

doi: 10.1038/ncomms11508

Inner ear organoids generated from Atoh1/nGFP ES cells include nGFP+ cells that expressed multiple hair cell markers. ( a , b ) The appearance of nGFP+ cells in the epithelium of organoid vesicles on day 12 of differentiation. ( c ) At later stages of differentiation ( > day 14), numerous nGFP+ cells populated the epithelium, typically in a regional cluster as seen in d and e . ( f , g ) Most nGFP+ cells were also Anx4a+ and Calretinin (Calb2)+. Arrowheads in g denote nGFP+ cells devoid of Calb2. ( h , i ) nGFP+ hair cells had acetylated-tublin+ kinocilia and F-actin+ hair bundles. ( j , k ) Most Calb2+ hair cells also had Espin+ hair bundles. ( l ) nGFP+ cells were also Myo7a+ and Sox2+. Scale bars, 100 μm ( a , c , d and e ), 25 μm ( b ) and 10 μm ( f – l ).
Figure Legend Snippet: Inner ear organoids generated from Atoh1/nGFP ES cells include nGFP+ cells that expressed multiple hair cell markers. ( a , b ) The appearance of nGFP+ cells in the epithelium of organoid vesicles on day 12 of differentiation. ( c ) At later stages of differentiation ( > day 14), numerous nGFP+ cells populated the epithelium, typically in a regional cluster as seen in d and e . ( f , g ) Most nGFP+ cells were also Anx4a+ and Calretinin (Calb2)+. Arrowheads in g denote nGFP+ cells devoid of Calb2. ( h , i ) nGFP+ hair cells had acetylated-tublin+ kinocilia and F-actin+ hair bundles. ( j , k ) Most Calb2+ hair cells also had Espin+ hair bundles. ( l ) nGFP+ cells were also Myo7a+ and Sox2+. Scale bars, 100 μm ( a , c , d and e ), 25 μm ( b ) and 10 μm ( f – l ).

Techniques Used: Generated

5) Product Images from "Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells"

Article Title: Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0087092

EGFP-positive cells exhibit a greater capacity for differentiation than EGFP-negative cells. (A) Sox2 expression was detected by IHC in xenograft tissues formed by SiHa-EGFP+, SiHa-EGFP−, C33A-EGFP+ and C33A-EGFP− cells. (B) The percentages of EGFP+ cells in the first, second, and third passages in differentiation medium. Bars = SE. *, p
Figure Legend Snippet: EGFP-positive cells exhibit a greater capacity for differentiation than EGFP-negative cells. (A) Sox2 expression was detected by IHC in xenograft tissues formed by SiHa-EGFP+, SiHa-EGFP−, C33A-EGFP+ and C33A-EGFP− cells. (B) The percentages of EGFP+ cells in the first, second, and third passages in differentiation medium. Bars = SE. *, p

Techniques Used: Expressing, Immunohistochemistry

Distinct molecular and biological properties of Sox2-positive and Sox2-negative cells. (A) Differential expression of several stem cell-related genes in SiHa-EGFP + and SiHa-EGFP − fractions validated by qPCR. (B) Detection of stem cell-related factors in sorted SiHa-EGFP + and SiHa-EGFP − cells by western blot. (C) Semi-quantitative analysis of stem cell-related factors relative to β-actin. (D) Stem cell-related gene expression in tumor xenografts was detected by immunohistochemistry. ALDH1 was detected by immunohistochemistry (E), western blot (F), and semi-quantitative analysis (G) in SiHa-EGFP + and SiHa-EGFP − tumors. β-actin was used as the loading control for RT-PCR and western blotting. Error bars represent S.D. (n = 3). * p
Figure Legend Snippet: Distinct molecular and biological properties of Sox2-positive and Sox2-negative cells. (A) Differential expression of several stem cell-related genes in SiHa-EGFP + and SiHa-EGFP − fractions validated by qPCR. (B) Detection of stem cell-related factors in sorted SiHa-EGFP + and SiHa-EGFP − cells by western blot. (C) Semi-quantitative analysis of stem cell-related factors relative to β-actin. (D) Stem cell-related gene expression in tumor xenografts was detected by immunohistochemistry. ALDH1 was detected by immunohistochemistry (E), western blot (F), and semi-quantitative analysis (G) in SiHa-EGFP + and SiHa-EGFP − tumors. β-actin was used as the loading control for RT-PCR and western blotting. Error bars represent S.D. (n = 3). * p

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction

Sorting living cervical cancer cells using the nuclear protein marker Sox2. Sox2 protein expression was detected by western blot (A) and immunohistochemistry (B) in the cervical cancer cell lines HeLa, SiHa, CaSki, and C33A. (C) The abundance of EGFP-positive tumor cells in cervical cancer cell lines transfected with the pSox2/EGFP reporter. (D) Immunohistochemical analysis of EGFP + and EGFP − cells. (E) Sox2 protein expression in a monolayer, tumorsphere cells, and sorted EGFP + and EGFP − cells.
Figure Legend Snippet: Sorting living cervical cancer cells using the nuclear protein marker Sox2. Sox2 protein expression was detected by western blot (A) and immunohistochemistry (B) in the cervical cancer cell lines HeLa, SiHa, CaSki, and C33A. (C) The abundance of EGFP-positive tumor cells in cervical cancer cell lines transfected with the pSox2/EGFP reporter. (D) Immunohistochemical analysis of EGFP + and EGFP − cells. (E) Sox2 protein expression in a monolayer, tumorsphere cells, and sorted EGFP + and EGFP − cells.

Techniques Used: Marker, Expressing, Western Blot, Immunohistochemistry, Transfection

Sox2-positive cells exhibit more EMT features but do not show altered proliferation in vitro . (A) Real-time PCR analysis of the mRNA levels of various EMT-related genes in SiHa-EGFP + and SiHa-EGFP − cells. EMT-related gene expression was measured by western blot (B) and immunochemistry (C) in SiHa-EGFP + and SiHa-EGFP − cells. (D) Immunochemistry for Sox2 and EMT-related genes in tumor xenografts. Bars = SE. *, p
Figure Legend Snippet: Sox2-positive cells exhibit more EMT features but do not show altered proliferation in vitro . (A) Real-time PCR analysis of the mRNA levels of various EMT-related genes in SiHa-EGFP + and SiHa-EGFP − cells. EMT-related gene expression was measured by western blot (B) and immunochemistry (C) in SiHa-EGFP + and SiHa-EGFP − cells. (D) Immunochemistry for Sox2 and EMT-related genes in tumor xenografts. Bars = SE. *, p

Techniques Used: In Vitro, Real-time Polymerase Chain Reaction, Expressing, Western Blot

6) Product Images from "Modulation of Wnt Signaling Enhances Inner Ear Organoid Development in 3D Culture"

Article Title: Modulation of Wnt Signaling Enhances Inner Ear Organoid Development in 3D Culture

Journal: PLoS ONE

doi: 10.1371/journal.pone.0162508

CHIR-treated aggregates give rise to inner ear organoids harboring mechanosensitive hair cells. (A-B) Co-localization of two hair cell markers Calb2 and Myo7a (A) or Sox2 and Myo7a (B) in cells lining the luminal surface of a vesicle. (C) Atoh1/nGFP + cells also express Myo7a and Sox2. (D-F) Cells expressing Brn3C, Myo7a, and Atoh1/nGFP exhibit flask-like morphology with a hair bundle on their apical surface characteristic of vestibular hair cells. Some Myo7a + cells in day 28 samples have faint Atoh1/nGFP expression (arrows), suggesting that these cells are more mature hair cells than cells expressing strong Atoh1/nGFP expression. (G) The hair bundle marker Espin was observed on the apical surface of Calb2 + hair cells. (H) Representative voltage-gated currents and mechanosensitive currents recorded from day 25 Atoh1/nGFP + cells in response to voltage injections and hair bundle deflections, respectively. (I) A cluster of Brn3A + neuronal cell bodies were located near Myo7a + hair cells. (J) A TUJ1 + neural processes extend and contact Myo7a + hair cells. (K) The ribbon synapse marker CtBP2 was associated with Myo7a + cells and TUJ1 + processes. Scale bars, 10 μm (A-G, I-K).
Figure Legend Snippet: CHIR-treated aggregates give rise to inner ear organoids harboring mechanosensitive hair cells. (A-B) Co-localization of two hair cell markers Calb2 and Myo7a (A) or Sox2 and Myo7a (B) in cells lining the luminal surface of a vesicle. (C) Atoh1/nGFP + cells also express Myo7a and Sox2. (D-F) Cells expressing Brn3C, Myo7a, and Atoh1/nGFP exhibit flask-like morphology with a hair bundle on their apical surface characteristic of vestibular hair cells. Some Myo7a + cells in day 28 samples have faint Atoh1/nGFP expression (arrows), suggesting that these cells are more mature hair cells than cells expressing strong Atoh1/nGFP expression. (G) The hair bundle marker Espin was observed on the apical surface of Calb2 + hair cells. (H) Representative voltage-gated currents and mechanosensitive currents recorded from day 25 Atoh1/nGFP + cells in response to voltage injections and hair bundle deflections, respectively. (I) A cluster of Brn3A + neuronal cell bodies were located near Myo7a + hair cells. (J) A TUJ1 + neural processes extend and contact Myo7a + hair cells. (K) The ribbon synapse marker CtBP2 was associated with Myo7a + cells and TUJ1 + processes. Scale bars, 10 μm (A-G, I-K).

Techniques Used: Expressing, Marker

CHIR treatment has dose-dependent effects on the number of vesicles containing Myo7a + /Sox2 + cells. (A-D’) Representative images show Myo7a/Sox2 expression in day 21 aggregate sensory epithelia that received 0 μM (A, A’), 1 μM (B, B’), 3 μM (C, C’), and 10 μM (D, D’) CHIR between days 8 and 10. (E) The average number of vesicles containing Myo7a + /Sox2 + cells per aggregate as a function of the CHIR concentration (**P
Figure Legend Snippet: CHIR treatment has dose-dependent effects on the number of vesicles containing Myo7a + /Sox2 + cells. (A-D’) Representative images show Myo7a/Sox2 expression in day 21 aggregate sensory epithelia that received 0 μM (A, A’), 1 μM (B, B’), 3 μM (C, C’), and 10 μM (D, D’) CHIR between days 8 and 10. (E) The average number of vesicles containing Myo7a + /Sox2 + cells per aggregate as a function of the CHIR concentration (**P

Techniques Used: Expressing, Concentration Assay

7) Product Images from "The secretome of periodontal ligament stem cells from MS patients protects against EAE"

Article Title: The secretome of periodontal ligament stem cells from MS patients protects against EAE

Journal: Scientific Reports

doi: 10.1038/srep38743

Flow cytometry of hPDLSCs and RR-MS-hPDLSCs phenotypes. Flow cytometry phenotype of surface related antigen of hPDLSCs and RR-MS-hPDLSCs at the 2 nd passage (CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD117, CD133, CD144, CD146, CD166, CD326, HLA-ABC, HLA-DR) and intracellular stemness (SSEA4, Oct3/4, Sox2, NANOG) marker expression levels were detected. Red histograms show the distribution of each antigen expression, whereas Blue histograms represent the distribution of the respective background control. Data are representative of five separate experiments.
Figure Legend Snippet: Flow cytometry of hPDLSCs and RR-MS-hPDLSCs phenotypes. Flow cytometry phenotype of surface related antigen of hPDLSCs and RR-MS-hPDLSCs at the 2 nd passage (CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD117, CD133, CD144, CD146, CD166, CD326, HLA-ABC, HLA-DR) and intracellular stemness (SSEA4, Oct3/4, Sox2, NANOG) marker expression levels were detected. Red histograms show the distribution of each antigen expression, whereas Blue histograms represent the distribution of the respective background control. Data are representative of five separate experiments.

Techniques Used: Flow Cytometry, Cytometry, Mass Spectrometry, Marker, Expressing

8) Product Images from "Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells"

Article Title: Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0087092

EGFP-positive cells exhibit a greater capacity for differentiation than EGFP-negative cells. (A) Sox2 expression was detected by IHC in xenograft tissues formed by SiHa-EGFP+, SiHa-EGFP−, C33A-EGFP+ and C33A-EGFP− cells. (B) The percentages of EGFP+ cells in the first, second, and third passages in differentiation medium. Bars = SE. *, p
Figure Legend Snippet: EGFP-positive cells exhibit a greater capacity for differentiation than EGFP-negative cells. (A) Sox2 expression was detected by IHC in xenograft tissues formed by SiHa-EGFP+, SiHa-EGFP−, C33A-EGFP+ and C33A-EGFP− cells. (B) The percentages of EGFP+ cells in the first, second, and third passages in differentiation medium. Bars = SE. *, p

Techniques Used: Expressing, Immunohistochemistry

Distinct molecular and biological properties of Sox2-positive and Sox2-negative cells. (A) Differential expression of several stem cell-related genes in SiHa-EGFP + and SiHa-EGFP − fractions validated by qPCR. (B) Detection of stem cell-related factors in sorted SiHa-EGFP + and SiHa-EGFP − cells by western blot. (C) Semi-quantitative analysis of stem cell-related factors relative to β-actin. (D) Stem cell-related gene expression in tumor xenografts was detected by immunohistochemistry. ALDH1 was detected by immunohistochemistry (E), western blot (F), and semi-quantitative analysis (G) in SiHa-EGFP + and SiHa-EGFP − tumors. β-actin was used as the loading control for RT-PCR and western blotting. Error bars represent S.D. (n = 3). * p
Figure Legend Snippet: Distinct molecular and biological properties of Sox2-positive and Sox2-negative cells. (A) Differential expression of several stem cell-related genes in SiHa-EGFP + and SiHa-EGFP − fractions validated by qPCR. (B) Detection of stem cell-related factors in sorted SiHa-EGFP + and SiHa-EGFP − cells by western blot. (C) Semi-quantitative analysis of stem cell-related factors relative to β-actin. (D) Stem cell-related gene expression in tumor xenografts was detected by immunohistochemistry. ALDH1 was detected by immunohistochemistry (E), western blot (F), and semi-quantitative analysis (G) in SiHa-EGFP + and SiHa-EGFP − tumors. β-actin was used as the loading control for RT-PCR and western blotting. Error bars represent S.D. (n = 3). * p

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction

Sorting living cervical cancer cells using the nuclear protein marker Sox2. Sox2 protein expression was detected by western blot (A) and immunohistochemistry (B) in the cervical cancer cell lines HeLa, SiHa, CaSki, and C33A. (C) The abundance of EGFP-positive tumor cells in cervical cancer cell lines transfected with the pSox2/EGFP reporter. (D) Immunohistochemical analysis of EGFP + and EGFP − cells. (E) Sox2 protein expression in a monolayer, tumorsphere cells, and sorted EGFP + and EGFP − cells.
Figure Legend Snippet: Sorting living cervical cancer cells using the nuclear protein marker Sox2. Sox2 protein expression was detected by western blot (A) and immunohistochemistry (B) in the cervical cancer cell lines HeLa, SiHa, CaSki, and C33A. (C) The abundance of EGFP-positive tumor cells in cervical cancer cell lines transfected with the pSox2/EGFP reporter. (D) Immunohistochemical analysis of EGFP + and EGFP − cells. (E) Sox2 protein expression in a monolayer, tumorsphere cells, and sorted EGFP + and EGFP − cells.

Techniques Used: Marker, Expressing, Western Blot, Immunohistochemistry, Transfection

Sox2-positive cells exhibit more EMT features but do not show altered proliferation in vitro . (A) Real-time PCR analysis of the mRNA levels of various EMT-related genes in SiHa-EGFP + and SiHa-EGFP − cells. EMT-related gene expression was measured by western blot (B) and immunochemistry (C) in SiHa-EGFP + and SiHa-EGFP − cells. (D) Immunochemistry for Sox2 and EMT-related genes in tumor xenografts. Bars = SE. *, p
Figure Legend Snippet: Sox2-positive cells exhibit more EMT features but do not show altered proliferation in vitro . (A) Real-time PCR analysis of the mRNA levels of various EMT-related genes in SiHa-EGFP + and SiHa-EGFP − cells. EMT-related gene expression was measured by western blot (B) and immunochemistry (C) in SiHa-EGFP + and SiHa-EGFP − cells. (D) Immunochemistry for Sox2 and EMT-related genes in tumor xenografts. Bars = SE. *, p

Techniques Used: In Vitro, Real-time Polymerase Chain Reaction, Expressing, Western Blot

9) Product Images from "Generation of inner ear sensory epithelia from pluripotent stem cells in 3D culture"

Article Title: Generation of inner ear sensory epithelia from pluripotent stem cells in 3D culture

Journal: Nature

doi: 10.1038/nature12298

Stem cell-derived otic vesicles generate functional inner ear hair cells a, b, Expression of Myo7a in the E9.5 otic vesicle ( a ; OtV) and day 14 vesicles ( b ). nb, neuroblasts. c-e, Myo7a/Sox2 + hair cells (hc) with underlying Sox2 + supporting cells (sc) on day 15 ( c ) and 16 ( d, e ). f-i, Whole-mount immunofluorescence for Myo7a and Sox2 ( f ) and 3D reconstruction ( g-i ) of a vesicle in a day 20 BMP/SB-FGF/LDN aggregate. j , Vesicles display the hallmarks of inner ear sensory epithelia. k - m, F-actin (F-act) labels cell-cell junctions on the luminal surface and stereocilia bundles. m , Acetylated-a-Tubulin (Tublin) labels kinocilium and the cuticular plate. n , Transmission electron micrograph of stereocilia bundles and kinocilium (arrow). o , Distribution of stereocilia and kinocilium heights on days 20 and 24 compared to adult mouse utricle, range indicated by gray boxes (n > 100 cells; ± max/min). p , Representative hair cell following 1 min FM1-43FX incubation, fixation and staining for F-actin. q , Representative epithelium preparation (inset) and hair cell during electrophysiological recordings. r , Representative voltage-current responses recorded from hair cells. The voltage protocol is shown at the top. s, Day 20 aggregate with Myo7a/Sox2 + vesicles. epi, epidermis (dashed outline). t , Number of hair cells on day 20 (n=12–16; mean ± s.e.m.). Scale bars, 250 μm ( f, s, q-inset ), 50 μm ( d, g, h ), 25 μm ( a-c, e, i, k, l ), 10 μm ( q ), 5 μm ( m, p ), 250 nm ( n ).
Figure Legend Snippet: Stem cell-derived otic vesicles generate functional inner ear hair cells a, b, Expression of Myo7a in the E9.5 otic vesicle ( a ; OtV) and day 14 vesicles ( b ). nb, neuroblasts. c-e, Myo7a/Sox2 + hair cells (hc) with underlying Sox2 + supporting cells (sc) on day 15 ( c ) and 16 ( d, e ). f-i, Whole-mount immunofluorescence for Myo7a and Sox2 ( f ) and 3D reconstruction ( g-i ) of a vesicle in a day 20 BMP/SB-FGF/LDN aggregate. j , Vesicles display the hallmarks of inner ear sensory epithelia. k - m, F-actin (F-act) labels cell-cell junctions on the luminal surface and stereocilia bundles. m , Acetylated-a-Tubulin (Tublin) labels kinocilium and the cuticular plate. n , Transmission electron micrograph of stereocilia bundles and kinocilium (arrow). o , Distribution of stereocilia and kinocilium heights on days 20 and 24 compared to adult mouse utricle, range indicated by gray boxes (n > 100 cells; ± max/min). p , Representative hair cell following 1 min FM1-43FX incubation, fixation and staining for F-actin. q , Representative epithelium preparation (inset) and hair cell during electrophysiological recordings. r , Representative voltage-current responses recorded from hair cells. The voltage protocol is shown at the top. s, Day 20 aggregate with Myo7a/Sox2 + vesicles. epi, epidermis (dashed outline). t , Number of hair cells on day 20 (n=12–16; mean ± s.e.m.). Scale bars, 250 μm ( f, s, q-inset ), 50 μm ( d, g, h ), 25 μm ( a-c, e, i, k, l ), 10 μm ( q ), 5 μm ( m, p ), 250 nm ( n ).

Techniques Used: Derivative Assay, Functional Assay, Expressing, Immunofluorescence, Activated Clotting Time Assay, Transmission Assay, Incubation, Staining

10) 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

11) Product Images from "Assessment of the Potential of CDK2 Inhibitor NU6140 to Influence the Expression of Pluripotency Markers NANOG, OCT4, and SOX2 in 2102Ep and H9 Cells"

Article Title: Assessment of the Potential of CDK2 Inhibitor NU6140 to Influence the Expression of Pluripotency Markers NANOG, OCT4, and SOX2 in 2102Ep and H9 Cells

Journal: International Journal of Cell Biology

doi: 10.1155/2014/280638

NU6140 treatment compared to nocodazole treatment affects differently the expression of pluripotency markers in 2102Ep carcinoma cells. Expression of pluripotency markers NANOG, OCT4, and SOX2 in nocodazole and NU6140 treated 2102Ep cells as detected by using flow cytometric assays. (a) Fixed and permeabilised (methanol permeabilisation) cells were stained with anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) Correlation between expression of CDK2 with NANOG and SOX2. Fixed and permeabilised (methanol permeabilisation) cells were stained with anti-CDK2 (Alexa Fluor 488 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. Results are shown as density plots.
Figure Legend Snippet: NU6140 treatment compared to nocodazole treatment affects differently the expression of pluripotency markers in 2102Ep carcinoma cells. Expression of pluripotency markers NANOG, OCT4, and SOX2 in nocodazole and NU6140 treated 2102Ep cells as detected by using flow cytometric assays. (a) Fixed and permeabilised (methanol permeabilisation) cells were stained with anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) Correlation between expression of CDK2 with NANOG and SOX2. Fixed and permeabilised (methanol permeabilisation) cells were stained with anti-CDK2 (Alexa Fluor 488 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. Results are shown as density plots.

Techniques Used: Expressing, Flow Cytometry, Staining

NU6140 treatment causes changes in the colony structure and differentiation potential of hES cells. hES cells were treated with NU6140 (10 μ M) for 24 h and then cultured in fresh mTeSR1 maintenance medium for a further 3 days with the medium changed daily. Embryoid bodies were formed from hES cells treated with NU6140 or from untreated hES cells. (a) Morphological changes in colony structure after NU6140 treatment and formation of EBs. (b) Fixed and permeabilised hES cells were stained with anti-OCT4 (Alexa Fluor 647 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp-Cy5.5) antibodies and with DAPI. Other hES cells were stained with anti-GATA4 (NorthernLights, NL-493 conjugate), anti-OTX-2 (NL-557), anti-HAND1 (NL-637) antibodies and with DAPI. (c) Differentiation into three germ cell layers as detected by flow cytometric assay. Fixed (1.6% PFA) and permeabilised cells from dissociated embryoid bodies (EBs) were stained with anti-SOX1 (NL-493), anti-Brachyury (NL-557), anti-SOX17 (NL-637), and anti-OCT4 (PerCp-Cy5.5) antibodies and with DAPI.
Figure Legend Snippet: NU6140 treatment causes changes in the colony structure and differentiation potential of hES cells. hES cells were treated with NU6140 (10 μ M) for 24 h and then cultured in fresh mTeSR1 maintenance medium for a further 3 days with the medium changed daily. Embryoid bodies were formed from hES cells treated with NU6140 or from untreated hES cells. (a) Morphological changes in colony structure after NU6140 treatment and formation of EBs. (b) Fixed and permeabilised hES cells were stained with anti-OCT4 (Alexa Fluor 647 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp-Cy5.5) antibodies and with DAPI. Other hES cells were stained with anti-GATA4 (NorthernLights, NL-493 conjugate), anti-OTX-2 (NL-557), anti-HAND1 (NL-637) antibodies and with DAPI. (c) Differentiation into three germ cell layers as detected by flow cytometric assay. Fixed (1.6% PFA) and permeabilised cells from dissociated embryoid bodies (EBs) were stained with anti-SOX1 (NL-493), anti-Brachyury (NL-557), anti-SOX17 (NL-637), and anti-OCT4 (PerCp-Cy5.5) antibodies and with DAPI.

Techniques Used: Cell Culture, Staining, Flow Cytometry

NU6140 treatment compared to nocodazole treatment affects differently the expression of pluripotency markers in hES cells. (a) Flow cytometric analysis of the expression of pluripotency markers NANOG, OCT4, and SOX2 in hES cells treated with nocodazole, NU6140, and DMSO. Fixed and permeabilised cells were stained with anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) Correlation between expression of CDK2 with NANOG and SOX2. Fixed and permeabilised cells were stained with anti-CDK2 (Alexa Fluor 488 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI.
Figure Legend Snippet: NU6140 treatment compared to nocodazole treatment affects differently the expression of pluripotency markers in hES cells. (a) Flow cytometric analysis of the expression of pluripotency markers NANOG, OCT4, and SOX2 in hES cells treated with nocodazole, NU6140, and DMSO. Fixed and permeabilised cells were stained with anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI. For analysis cellular debris and doublets were excluded. (b) Correlation between expression of CDK2 with NANOG and SOX2. Fixed and permeabilised cells were stained with anti-CDK2 (Alexa Fluor 488 conjugate), anti-NANOG (PE), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies and with DAPI.

Techniques Used: Expressing, Flow Cytometry, Staining

Effect of combined treatment on pluripotency markers expression in hES and hEC cells compared to treatments with nocodazole or NU6140 individually. Nocodazole treated cells (10 h) were washed and further treated with 10 μ M NU6140 for 14 h. The coexpression of NANOG/OCT4 and SOX2/OCT4 detected in hEC (a, c) as described in Figure 2 and in hES cells (b, d) as described in Figure 1 . (e) Morphological changes in colony structure of hES cells treated with NU6140 or nocodazole.
Figure Legend Snippet: Effect of combined treatment on pluripotency markers expression in hES and hEC cells compared to treatments with nocodazole or NU6140 individually. Nocodazole treated cells (10 h) were washed and further treated with 10 μ M NU6140 for 14 h. The coexpression of NANOG/OCT4 and SOX2/OCT4 detected in hEC (a, c) as described in Figure 2 and in hES cells (b, d) as described in Figure 1 . (e) Morphological changes in colony structure of hES cells treated with NU6140 or nocodazole.

Techniques Used: Expressing

12) Product Images from "SOX2 Is Regulated Differently from NANOG and OCT4 in Human Embryonic Stem Cells during Early Differentiation Initiated with Sodium Butyrate"

Article Title: SOX2 Is Regulated Differently from NANOG and OCT4 in Human Embryonic Stem Cells during Early Differentiation Initiated with Sodium Butyrate

Journal: Stem Cells International

doi: 10.1155/2014/298163

Detection of pluripotency marker SOX2 and differentiation markers in differentiating H9 cells. Cells were treated and processed as described in the legend of Figure 2 . Coexpression of pluripotency marker SOX2 and differentiation markers was detected by flow cytometry (a) or by Western blot (b). Results are representative of two independent experiments. (c) The number of cells in one well is shown as a mean value ± SEM of three experiments. (d) The changes in colony morphology detected at the beginning of differentiation (day 0) and on day 3.
Figure Legend Snippet: Detection of pluripotency marker SOX2 and differentiation markers in differentiating H9 cells. Cells were treated and processed as described in the legend of Figure 2 . Coexpression of pluripotency marker SOX2 and differentiation markers was detected by flow cytometry (a) or by Western blot (b). Results are representative of two independent experiments. (c) The number of cells in one well is shown as a mean value ± SEM of three experiments. (d) The changes in colony morphology detected at the beginning of differentiation (day 0) and on day 3.

Techniques Used: Marker, Flow Cytometry, Cytometry, Western Blot

Characterisation of SOX17 and GATA4 expressing cells for expression of pluripotency markers NANOG, OCT4, and SOX2. Cells coexpressing SOX17 (a) or GATA-4 (b) and SOX2 were selected and coexpression of NANOG and OCT4 in these cells was estimated.
Figure Legend Snippet: Characterisation of SOX17 and GATA4 expressing cells for expression of pluripotency markers NANOG, OCT4, and SOX2. Cells coexpressing SOX17 (a) or GATA-4 (b) and SOX2 were selected and coexpression of NANOG and OCT4 in these cells was estimated.

Techniques Used: Expressing

Changes in the expression patterns of pluripotency markers NANOG and OCT4 are distinct from those of SOX2 in differentiating H9 cells. hES cells were treated with sodium butyrate as described in Section 2 . Coexpression of pluripotency markers during differentiation on days 3 and 4 was detected by flow cytometry. Fixed and permeabilised cells were stained with anti-SSEA-3 (Alexa Fluor 488 conjugate), anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies as well as with DAPI. Cellular debris and doublets were excluded from analysis. Results are representative of two independent experiments.
Figure Legend Snippet: Changes in the expression patterns of pluripotency markers NANOG and OCT4 are distinct from those of SOX2 in differentiating H9 cells. hES cells were treated with sodium butyrate as described in Section 2 . Coexpression of pluripotency markers during differentiation on days 3 and 4 was detected by flow cytometry. Fixed and permeabilised cells were stained with anti-SSEA-3 (Alexa Fluor 488 conjugate), anti-NANOG (PE), anti-OCT4 (Alexa Fluor 647), and anti-SOX2 (PerCp Cy5.5 conjugate) antibodies as well as with DAPI. Cellular debris and doublets were excluded from analysis. Results are representative of two independent experiments.

Techniques Used: Expressing, Flow Cytometry, Cytometry, Staining

13) Product Images from "Efficient Generation of Integration-Free iPS Cells from Human Adult Peripheral Blood Using BCL-XL Together with Yamanaka Factors"

Article Title: Efficient Generation of Integration-Free iPS Cells from Human Adult Peripheral Blood Using BCL-XL Together with Yamanaka Factors

Journal: PLoS ONE

doi: 10.1371/journal.pone.0064496

Generation of integration-free iPSCs from adult PBMNCs with episomal vectors. ( A ) ALP staining of iPSCs at 4 weeks after nucleofection of PBMNCs with reprogramming factor-expressing episomal vectors. OS, OCT4 and SOX2; MK, MYC and KLF4; B, BCL-XL. PBMNCs were cultured for 4–8 days before nucleofection. 1×10 6 PBMNCs were nucleofected and then seeded into each well. ( B ) Inclusion of BCL-XL increases PB reprogramming efficiency by up to 10-fold. PBMNCs were cultured for 4–8 days before nucleofection. ALP-positive iPSC colonies were enumerated at 3–4 weeks after nucleofection. Data are presented as mean ± SEM (n = 6). In all 3 conditions, BCL-XL significantly increased reprogramming efficiency. * indicates P
Figure Legend Snippet: Generation of integration-free iPSCs from adult PBMNCs with episomal vectors. ( A ) ALP staining of iPSCs at 4 weeks after nucleofection of PBMNCs with reprogramming factor-expressing episomal vectors. OS, OCT4 and SOX2; MK, MYC and KLF4; B, BCL-XL. PBMNCs were cultured for 4–8 days before nucleofection. 1×10 6 PBMNCs were nucleofected and then seeded into each well. ( B ) Inclusion of BCL-XL increases PB reprogramming efficiency by up to 10-fold. PBMNCs were cultured for 4–8 days before nucleofection. ALP-positive iPSC colonies were enumerated at 3–4 weeks after nucleofection. Data are presented as mean ± SEM (n = 6). In all 3 conditions, BCL-XL significantly increased reprogramming efficiency. * indicates P

Techniques Used: ALP Assay, Staining, Expressing, Cell Culture

Characterization of integration-free PB iPSCs. ( A ) PB iPSCs show a typical morphology of human pluripotent stem cells. ( B ) A representative karyogram of an iPSC clone. All analyzed PB iPSC clones showed a normal karyotype. ( C ) PB iPSCs express pluripotency markers OCT4, SOX2, NANOG and SSEA4. Shown are representative confocal images captured using the Zeiss LSM 710 confocal microscope with a 20× objective. ( D ) PB iPSCs form teratoma in immunodeficient mice. H E staining of representative teratoma from PB iPSCs shows derivatives of 3 embryonic germ layers. Cartilage (mesoderm); glands (endoderm) and neurotubules (ectoderm). Images were acquired using the Olympus microscope with a 20× objective.
Figure Legend Snippet: Characterization of integration-free PB iPSCs. ( A ) PB iPSCs show a typical morphology of human pluripotent stem cells. ( B ) A representative karyogram of an iPSC clone. All analyzed PB iPSC clones showed a normal karyotype. ( C ) PB iPSCs express pluripotency markers OCT4, SOX2, NANOG and SSEA4. Shown are representative confocal images captured using the Zeiss LSM 710 confocal microscope with a 20× objective. ( D ) PB iPSCs form teratoma in immunodeficient mice. H E staining of representative teratoma from PB iPSCs shows derivatives of 3 embryonic germ layers. Cartilage (mesoderm); glands (endoderm) and neurotubules (ectoderm). Images were acquired using the Olympus microscope with a 20× objective.

Techniques Used: Clone Assay, Microscopy, Mouse Assay, Staining

BCL-XL significantly enhances OS-mediated reprogramming of cord blood and peripheral blood cells with lentiviral vectors. ( A ) Differential effects of BCL2 family members on enhancing OS-mediated reprogramming of CB cells. CB CD34 + cells were cultured for 2 days before lentiviral transduction. CB iPSC colonies were enumerated at 2 weeks after transduction of reprogramming factors. Data shown are presented as mean ± SEM (n = 4). OS: OCT4 and SOX2. * indicates P
Figure Legend Snippet: BCL-XL significantly enhances OS-mediated reprogramming of cord blood and peripheral blood cells with lentiviral vectors. ( A ) Differential effects of BCL2 family members on enhancing OS-mediated reprogramming of CB cells. CB CD34 + cells were cultured for 2 days before lentiviral transduction. CB iPSC colonies were enumerated at 2 weeks after transduction of reprogramming factors. Data shown are presented as mean ± SEM (n = 4). OS: OCT4 and SOX2. * indicates P

Techniques Used: Cell Culture, Transduction

Related Articles

Western Blot:

Article Title: Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells
Article Snippet: .. After blocking with 5% fat-free milk in Tris-buffered saline, the following antibodies were used for western blotting: anti-Sox2 (1∶500), anti-ALDH1 (BD Biosciences, 1∶500), anti-Bmi1 (1∶500), anti-Oct4 (1∶500), anti-Nanog (1∶500), anti-vimentin (1∶500), anti-snail (1∶500), anti-β-catenin (1∶500), anti-E-cadherin (1∶500), and anti-β-actin (1∶1000) overnight at 4°C. ..

Flow Cytometry:

Article Title: A Conserved Mechanism for Control of Human and Mouse Embryonic Stem Cell Pluripotency and Differentiation by Shp2 Tyrosine Phosphatase
Article Snippet: .. Flow cytometry was performed using BD FACSCanto analyzer with Alexa647-conjugated antibodies against Ssea-1 (Santa Cruz), Oct3/4, Sox2 (from BD pharmagen). .. Transfection and in vitro differentiation of hESCs H14 cells were cultured 3–4 days before transfection.

Cytometry:

Article Title: A Conserved Mechanism for Control of Human and Mouse Embryonic Stem Cell Pluripotency and Differentiation by Shp2 Tyrosine Phosphatase
Article Snippet: .. Flow cytometry was performed using BD FACSCanto analyzer with Alexa647-conjugated antibodies against Ssea-1 (Santa Cruz), Oct3/4, Sox2 (from BD pharmagen). .. Transfection and in vitro differentiation of hESCs H14 cells were cultured 3–4 days before transfection.

Blocking Assay:

Article Title: Cervical Cancer Cells with Positive Sox2 Expression Exhibit the Properties of Cancer Stem Cells
Article Snippet: .. After blocking with 5% fat-free milk in Tris-buffered saline, the following antibodies were used for western blotting: anti-Sox2 (1∶500), anti-ALDH1 (BD Biosciences, 1∶500), anti-Bmi1 (1∶500), anti-Oct4 (1∶500), anti-Nanog (1∶500), anti-vimentin (1∶500), anti-snail (1∶500), anti-β-catenin (1∶500), anti-E-cadherin (1∶500), and anti-β-actin (1∶1000) overnight at 4°C. ..

Staining:

Article Title: Efficient Reprogramming of Human Cord Blood CD34+ Cells Into Induced Pluripotent Stem Cells With OCT4 and SOX2 Alone
Article Snippet: .. Cells were treated with fixation buffer and permeabilization buffer (eBiosciences) for 30 minutes before being stained overnight with PE or FITC-conjugated antibodies OCT4 (eBiosciences), SOX2 (BD Pharmingen), NANOG (BD Pharmingen), SSEA-3 (eBiosciences), SSEA-4 (eBiosciences), and TRA-1-60 (Stemgent). ..

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  • 86
    Becton Dickinson sox2 anti human
    Expression of OCT4 in combination with neural progenitor culture induces conversion of fibroblasts to a proliferating neural progenitor-like cell (hFib-NPC OCT4 ). (A) Schematic of conversion strategy. (B) Flow cytometric plot of intracellular OCT4 expression in adult human fibroblasts after 8 days of culture in reprogramming media culture (hFibR OCT4 ). (C) Phase-contrast image of hFibR OCT4 neural sphere-like clusters after 14 days in neural progenitor culture. (D) Flow cytometric plot of OCT4 versus Ki67 expression in hFibR OCT4 sphere-derived cells. (E) Flow cytometric plot of OCT4 versus <t>SOX2</t> expression in hFibR OCT4 sphere-derived cells. (F) Flow cytometric plot of eGFP expression from hFibR OCT4 cells generated with pHIV-ef1α-OCT4-IRES-eGFP versus CD133 on live cells. (G) Phase-contrast image of hFibR OCT4 sphere-derived cells cultured on poly-ornathine/laminin (POL). (H) Immunofluorescence results for OCT4 and SOX2 expression in hFibR OCT4 sphere-derived cells cultured on POL. (I) Cell growth curve illustrating cumulative number of viable trypan blue excluding cells with increasing passage number. (J) Flow cytometric plot of SOX2 versus Ki67 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (K) Flow cytometric plot of OCT4 versus SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (L) Immunofluorescence results for NESTIN and SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. Representative results from n =4 rounds of hFib-NPC OCT4 conversion. Scale bar=100 μM. White arrows denote areas of interest.
    Sox2 Anti Human, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti sox2
    Flow cytometry of hPDLSCs and RR-MS-hPDLSCs phenotypes. Flow cytometry phenotype of surface related antigen of hPDLSCs and RR-MS-hPDLSCs at the 2 nd passage (CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD117, CD133, CD144, CD146, CD166, CD326, HLA-ABC, HLA-DR) and intracellular stemness (SSEA4, Oct3/4, <t>Sox2,</t> NANOG) marker expression levels were detected. Red histograms show the distribution of each antigen expression, whereas Blue histograms represent the distribution of the respective background control. Data are representative of five separate experiments.
    Anti Sox2, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti sox2/product/Becton Dickinson
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    Becton Dickinson antibody nestin
    MiR-20 regulated NPCs differentiation. ( A ) qPCR data showing mRNA levels of <t>Nestin,</t> <t>Sox2,</t> Vimentin, Tuj1 and Map2 genes during NPCs differentiation. ( B–F ) Immunostaining images and quantified data of Nestin ( B ), Sox2 ( C ), Vimentin ( D ), Tuj1 ( E ) and Map2 ( F ) positive cells in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. Scale bar, 50 μm (Top panel: immunostaining images; Bottom panel: quantified data from positive immunostaining cells). Quantitation and representative photomicrographs showed that miR-20 promotes cell differentiation in NPCs. Bars show mean ± SD. All experiments were repeated three times. *P
    Antibody Nestin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson percp cy 5 5 conjugated anti sox2 17
    MiR-20 regulated NPCs differentiation. ( A ) qPCR data showing mRNA levels of <t>Nestin,</t> <t>Sox2,</t> Vimentin, Tuj1 and Map2 genes during NPCs differentiation. ( B–F ) Immunostaining images and quantified data of Nestin ( B ), Sox2 ( C ), Vimentin ( D ), Tuj1 ( E ) and Map2 ( F ) positive cells in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. Scale bar, 50 μm (Top panel: immunostaining images; Bottom panel: quantified data from positive immunostaining cells). Quantitation and representative photomicrographs showed that miR-20 promotes cell differentiation in NPCs. Bars show mean ± SD. All experiments were repeated three times. *P
    Percp Cy 5 5 Conjugated Anti Sox2 17, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression of OCT4 in combination with neural progenitor culture induces conversion of fibroblasts to a proliferating neural progenitor-like cell (hFib-NPC OCT4 ). (A) Schematic of conversion strategy. (B) Flow cytometric plot of intracellular OCT4 expression in adult human fibroblasts after 8 days of culture in reprogramming media culture (hFibR OCT4 ). (C) Phase-contrast image of hFibR OCT4 neural sphere-like clusters after 14 days in neural progenitor culture. (D) Flow cytometric plot of OCT4 versus Ki67 expression in hFibR OCT4 sphere-derived cells. (E) Flow cytometric plot of OCT4 versus SOX2 expression in hFibR OCT4 sphere-derived cells. (F) Flow cytometric plot of eGFP expression from hFibR OCT4 cells generated with pHIV-ef1α-OCT4-IRES-eGFP versus CD133 on live cells. (G) Phase-contrast image of hFibR OCT4 sphere-derived cells cultured on poly-ornathine/laminin (POL). (H) Immunofluorescence results for OCT4 and SOX2 expression in hFibR OCT4 sphere-derived cells cultured on POL. (I) Cell growth curve illustrating cumulative number of viable trypan blue excluding cells with increasing passage number. (J) Flow cytometric plot of SOX2 versus Ki67 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (K) Flow cytometric plot of OCT4 versus SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (L) Immunofluorescence results for NESTIN and SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. Representative results from n =4 rounds of hFib-NPC OCT4 conversion. Scale bar=100 μM. White arrows denote areas of interest.

    Journal: Stem Cells and Development

    Article Title: Activation of Neural Cell Fate Programs Toward Direct Conversion of Adult Human Fibroblasts into Tri-Potent Neural Progenitors Using OCT-4

    doi: 10.1089/scd.2014.0023

    Figure Lengend Snippet: Expression of OCT4 in combination with neural progenitor culture induces conversion of fibroblasts to a proliferating neural progenitor-like cell (hFib-NPC OCT4 ). (A) Schematic of conversion strategy. (B) Flow cytometric plot of intracellular OCT4 expression in adult human fibroblasts after 8 days of culture in reprogramming media culture (hFibR OCT4 ). (C) Phase-contrast image of hFibR OCT4 neural sphere-like clusters after 14 days in neural progenitor culture. (D) Flow cytometric plot of OCT4 versus Ki67 expression in hFibR OCT4 sphere-derived cells. (E) Flow cytometric plot of OCT4 versus SOX2 expression in hFibR OCT4 sphere-derived cells. (F) Flow cytometric plot of eGFP expression from hFibR OCT4 cells generated with pHIV-ef1α-OCT4-IRES-eGFP versus CD133 on live cells. (G) Phase-contrast image of hFibR OCT4 sphere-derived cells cultured on poly-ornathine/laminin (POL). (H) Immunofluorescence results for OCT4 and SOX2 expression in hFibR OCT4 sphere-derived cells cultured on POL. (I) Cell growth curve illustrating cumulative number of viable trypan blue excluding cells with increasing passage number. (J) Flow cytometric plot of SOX2 versus Ki67 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (K) Flow cytometric plot of OCT4 versus SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. (L) Immunofluorescence results for NESTIN and SOX2 expression in passage 5 POL cultured hFibR OCT4 sphere-derived cells. Representative results from n =4 rounds of hFib-NPC OCT4 conversion. Scale bar=100 μM. White arrows denote areas of interest.

    Article Snippet: Neural lineage cells were identified by staining with monoclonal antibodies OCT4-PE (BD), SOX2-647 (BD), SOX2 anti-human (BD), Nestin anti-Human (R & D), Beta-III Tubulin/Tuj1 anti-Human (R & D), Microtubule-associated protein 2/MAP2 anti-Human (Abcam), Oligodendrocyte marker 4/O4 anti-Human (R & D), Glial fibrilliary acidic protein/GFAP anti-Human (SIGMA), and CD133-PE (Miltenyi).

    Techniques: Expressing, Flow Cytometry, Derivative Assay, Generated, Cell Culture, Immunofluorescence

    Expression of SOX2 in combination with neural progenitor culture fails to induce conversion of adult fibroblasts to neural progenitors, despite successfully activating downstream neural conversion transcription factors. (A) Flow cytometric plot of intracellular SOX2 expression in adult human fibroblasts after 8 days in reprogramming media (hFibR SOX2 ). (B) Phase-contrast image of hFibR SOX2 sphere-like clusters after 14 days in neural progenitor culture. (C) Flow cytometric plot of SOX2 versus OCT4 expression in hFibR SOX2 sphere-derived cells. (D) Flow cytometric plot of SOX2 versus Ki67 expression in hFibR SOX2 sphere-derived cells. (E) Phase-contrast image of hFibR SOX2 sphere-derived cells cultured on POL. (F) Cell growth curve illustrating cumulative number of viable trypan blue excluding cells with increasing passage number. (G–J) qRT-PCR results for expression of neural conversion transcription factors in hFib eGFP cultured in neural progenitor culture, and hFibR SOX2 cultured in neural progenitor culture. Expression levels are normalized to expression in hFibs ( n =2). Representative results from n =3 attempts of hFibR SOX2 conversion. White arrows indicate areas of interest.

    Journal: Stem Cells and Development

    Article Title: Activation of Neural Cell Fate Programs Toward Direct Conversion of Adult Human Fibroblasts into Tri-Potent Neural Progenitors Using OCT-4

    doi: 10.1089/scd.2014.0023

    Figure Lengend Snippet: Expression of SOX2 in combination with neural progenitor culture fails to induce conversion of adult fibroblasts to neural progenitors, despite successfully activating downstream neural conversion transcription factors. (A) Flow cytometric plot of intracellular SOX2 expression in adult human fibroblasts after 8 days in reprogramming media (hFibR SOX2 ). (B) Phase-contrast image of hFibR SOX2 sphere-like clusters after 14 days in neural progenitor culture. (C) Flow cytometric plot of SOX2 versus OCT4 expression in hFibR SOX2 sphere-derived cells. (D) Flow cytometric plot of SOX2 versus Ki67 expression in hFibR SOX2 sphere-derived cells. (E) Phase-contrast image of hFibR SOX2 sphere-derived cells cultured on POL. (F) Cell growth curve illustrating cumulative number of viable trypan blue excluding cells with increasing passage number. (G–J) qRT-PCR results for expression of neural conversion transcription factors in hFib eGFP cultured in neural progenitor culture, and hFibR SOX2 cultured in neural progenitor culture. Expression levels are normalized to expression in hFibs ( n =2). Representative results from n =3 attempts of hFibR SOX2 conversion. White arrows indicate areas of interest.

    Article Snippet: Neural lineage cells were identified by staining with monoclonal antibodies OCT4-PE (BD), SOX2-647 (BD), SOX2 anti-human (BD), Nestin anti-Human (R & D), Beta-III Tubulin/Tuj1 anti-Human (R & D), Microtubule-associated protein 2/MAP2 anti-Human (Abcam), Oligodendrocyte marker 4/O4 anti-Human (R & D), Glial fibrilliary acidic protein/GFAP anti-Human (SIGMA), and CD133-PE (Miltenyi).

    Techniques: Expressing, Flow Cytometry, Derivative Assay, Cell Culture, Quantitative RT-PCR

    Flow cytometry of hPDLSCs and RR-MS-hPDLSCs phenotypes. Flow cytometry phenotype of surface related antigen of hPDLSCs and RR-MS-hPDLSCs at the 2 nd passage (CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD117, CD133, CD144, CD146, CD166, CD326, HLA-ABC, HLA-DR) and intracellular stemness (SSEA4, Oct3/4, Sox2, NANOG) marker expression levels were detected. Red histograms show the distribution of each antigen expression, whereas Blue histograms represent the distribution of the respective background control. Data are representative of five separate experiments.

    Journal: Scientific Reports

    Article Title: The secretome of periodontal ligament stem cells from MS patients protects against EAE

    doi: 10.1038/srep38743

    Figure Lengend Snippet: Flow cytometry of hPDLSCs and RR-MS-hPDLSCs phenotypes. Flow cytometry phenotype of surface related antigen of hPDLSCs and RR-MS-hPDLSCs at the 2 nd passage (CD13, CD14, CD29, CD31, CD34, CD44, CD45, CD73, CD90, CD105, CD106, CD117, CD133, CD144, CD146, CD166, CD326, HLA-ABC, HLA-DR) and intracellular stemness (SSEA4, Oct3/4, Sox2, NANOG) marker expression levels were detected. Red histograms show the distribution of each antigen expression, whereas Blue histograms represent the distribution of the respective background control. Data are representative of five separate experiments.

    Article Snippet: Cells were stained using the following antibodies: anti-CD13, anti-CD29, anti-CD44, anti-CD45, anti-CD105, anti-CD166 (Ancell, MN, USA), anti-CD14, anti-CD133 (BergischGladbach, Germany),anti-CD63, anti-CD73, anti-CD90, anti-CD117, anti-CD146, anti-CD271, anti-Sox2, anti-HLA-DR, anti-SSEA4, anti-OCT3/4 (Becton Dickinson, BD, San Jose, CA, USA); anti-CD144 (Acris Antibodies, Herford, Germany), anti-CD34 (Beckman Coulter, Fullerton, CA, USA).

    Techniques: Flow Cytometry, Cytometry, Mass Spectrometry, Marker, Expressing

    MiR-20 regulated NPCs differentiation. ( A ) qPCR data showing mRNA levels of Nestin, Sox2, Vimentin, Tuj1 and Map2 genes during NPCs differentiation. ( B–F ) Immunostaining images and quantified data of Nestin ( B ), Sox2 ( C ), Vimentin ( D ), Tuj1 ( E ) and Map2 ( F ) positive cells in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. Scale bar, 50 μm (Top panel: immunostaining images; Bottom panel: quantified data from positive immunostaining cells). Quantitation and representative photomicrographs showed that miR-20 promotes cell differentiation in NPCs. Bars show mean ± SD. All experiments were repeated three times. *P

    Journal: Scientific Reports

    Article Title: The miR-20-Rest-Wnt signaling axis regulates neural progenitor cell differentiation

    doi: 10.1038/srep23300

    Figure Lengend Snippet: MiR-20 regulated NPCs differentiation. ( A ) qPCR data showing mRNA levels of Nestin, Sox2, Vimentin, Tuj1 and Map2 genes during NPCs differentiation. ( B–F ) Immunostaining images and quantified data of Nestin ( B ), Sox2 ( C ), Vimentin ( D ), Tuj1 ( E ) and Map2 ( F ) positive cells in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. Scale bar, 50 μm (Top panel: immunostaining images; Bottom panel: quantified data from positive immunostaining cells). Quantitation and representative photomicrographs showed that miR-20 promotes cell differentiation in NPCs. Bars show mean ± SD. All experiments were repeated three times. *P

    Article Snippet: The cells were permeabilized with BD Phosflow™ Perm Buffer I (Cat. No. 557885), and then stained with antibody Nestin (561231, Becton, Dickinson and Company;), Sox2 (ab75485, Cambridge, USA), Vimentin (ab128507, Cambridge, USA), Tuj1(ab195879, Cambridge, USA), Map2 (560399, Becton, Dickinson and Company), GFAP (ab4674, Cambridge, USA) and theire Isotype control (550795, Becton, Dickinson and Company; ab170190, Cambridge, USA; ab91356, Cambridge, USA; ab171464 Cambridge, USA; 557721, Becton, Dickinson and Company; ab37382, Cambridge, USA).

    Techniques: Real-time Polymerase Chain Reaction, Immunostaining, Transfection, Quantitation Assay, Cell Differentiation

    The percentage of Nestin, Sox2, Vimentin, Tuj1, Map2 and GFAP positive cells determined by Fluorescence-activated sorting (FACS) analysis. Representative images showed the expression level of these genes in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. An isotype control is needed to determine whether fluorescence emitted is due to non-specific binding of the fluorescent antibody. The datas are shown as the means ± SD. From 3 independent repetitions. *P

    Journal: Scientific Reports

    Article Title: The miR-20-Rest-Wnt signaling axis regulates neural progenitor cell differentiation

    doi: 10.1038/srep23300

    Figure Lengend Snippet: The percentage of Nestin, Sox2, Vimentin, Tuj1, Map2 and GFAP positive cells determined by Fluorescence-activated sorting (FACS) analysis. Representative images showed the expression level of these genes in NPCs transfected with miRNA mimics, miRNA inhibitor or Rest siRNA alone in differentiation medium or differentiation medium containing Wnt3a or DKK1 for 96 h. An isotype control is needed to determine whether fluorescence emitted is due to non-specific binding of the fluorescent antibody. The datas are shown as the means ± SD. From 3 independent repetitions. *P

    Article Snippet: The cells were permeabilized with BD Phosflow™ Perm Buffer I (Cat. No. 557885), and then stained with antibody Nestin (561231, Becton, Dickinson and Company;), Sox2 (ab75485, Cambridge, USA), Vimentin (ab128507, Cambridge, USA), Tuj1(ab195879, Cambridge, USA), Map2 (560399, Becton, Dickinson and Company), GFAP (ab4674, Cambridge, USA) and theire Isotype control (550795, Becton, Dickinson and Company; ab170190, Cambridge, USA; ab91356, Cambridge, USA; ab171464 Cambridge, USA; 557721, Becton, Dickinson and Company; ab37382, Cambridge, USA).

    Techniques: Fluorescence, FACS, Expressing, Transfection, Binding Assay