Review



mixed rabbit anti oct1 antibodies  (Bethyl)


Bioz Verified Symbol Bethyl is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 93
      Buy from Supplier

    Structured Review

    Bethyl mixed rabbit anti oct1 antibodies
    A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse <t>anti-Oct1</t> antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.
    Mixed Rabbit Anti Oct1 Antibodies, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mixed+rabbit+anti+oct1+antibodies/pmc03493455-226-6-10?v=Bethyl
    Average 93 stars, based on 16 article reviews
    mixed rabbit anti oct1 antibodies - by Bioz Stars, 2026-07
    93/100 stars

    Images

    1) Product Images from "Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant"

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1003048

    A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.
    Figure Legend Snippet: A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.

    Techniques Used: Staining

    A. IF images are shown. Frozen malignant breast carcinoma sections (non-familial stage IIIA node-positive infiltrating ductal carcinoma, ER + PR + HER2 − ) were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and rabbit anti-ALDH1 antibodies (Abcam ab52492). Arrow indicates example double-positive cell. Asterisks show examples of an Oct1 HI cell with low ALDH1 and ALDH1 HI with low Oct1. Detail is shown in images below. B. Western blot depicting Oct1 levels in a panel of malignant epithelial cells isolated from pleural effusions (human breast carcinoma lung metastases). GAPDH is shown as a loading control. C. Examples of the CD24/44 profile from pleural effusions with highest and lowest Oct1 protein levels. D. Correlation of Oct1 protein and CD24 LO CD44 HI stem content in 15 individual patient samples (7 ER + PR + HER2 − , 5 ER − PR − HER2 − , 3 ER − PR − HER2 + ) collected from 14 different patients. One patient had tumor cells collected twice 22 months apart. Samples were placed into Oct1 LO and Oct1 HI categories based on Oct1 Western blot signal intensity relative to GAPDH staining. Cells were tested for percentage CD24 LO CD44 HI stem cell content and plotted. P -value was calculated using the two-tailed student T-test. E. The same analysis for Oct1 mRNA as measured by qRT-PCR.
    Figure Legend Snippet: A. IF images are shown. Frozen malignant breast carcinoma sections (non-familial stage IIIA node-positive infiltrating ductal carcinoma, ER + PR + HER2 − ) were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and rabbit anti-ALDH1 antibodies (Abcam ab52492). Arrow indicates example double-positive cell. Asterisks show examples of an Oct1 HI cell with low ALDH1 and ALDH1 HI with low Oct1. Detail is shown in images below. B. Western blot depicting Oct1 levels in a panel of malignant epithelial cells isolated from pleural effusions (human breast carcinoma lung metastases). GAPDH is shown as a loading control. C. Examples of the CD24/44 profile from pleural effusions with highest and lowest Oct1 protein levels. D. Correlation of Oct1 protein and CD24 LO CD44 HI stem content in 15 individual patient samples (7 ER + PR + HER2 − , 5 ER − PR − HER2 − , 3 ER − PR − HER2 + ) collected from 14 different patients. One patient had tumor cells collected twice 22 months apart. Samples were placed into Oct1 LO and Oct1 HI categories based on Oct1 Western blot signal intensity relative to GAPDH staining. Cells were tested for percentage CD24 LO CD44 HI stem cell content and plotted. P -value was calculated using the two-tailed student T-test. E. The same analysis for Oct1 mRNA as measured by qRT-PCR.

    Techniques Used: Staining, Western Blot, Isolation, Control, Two Tailed Test, Quantitative RT-PCR

    A. Aldefluor staining profile of A549 cells infected with a doxycycline-inducible lentiviral shRNA. MFI was 169.0 for scrambled and 91.2 for Oct1-specific shRNA. A549 cells were infected with control or Oct1-specific lentiviral particles (Santa Cruz), selected with puromycin, and subjected to analysis after 48 hr. B. Efficacy of the A549 knockdown as assessed by Western blotting using anti-Oct1 antibodies and an anti-GAPDH loading control. C. Oct1 Western blot of unsorted normal A549 cells cultured under normal conditions, and sorted Aldefluor HI and Aldefluor LO populations is shown. GAPDH is used as a loading control. D. The same sorted cells or unsorted cells were subjected to qRT-PCR to determine Oct1 mRNA levels. Levels are show relative to GAPDH. Error bars depict standard deviations. E. Oct1 was ectopically expressed in A549 cells using a retrovirus (pBabe-Oct1) or empty vector. The mixed population of cells was subjected to selection with puromycin, and ALDH activity determined. F. Western blot using anti-Oct1 antibodies of the same cells shown in (E). ß-actin is shown as a loading control. G. Alignment of the Aldh1a1 promoter regions in several example vertebrate species. The conserved perfect octamer sequence centered at approximately −55 bp relative to the transcription start site is highlighted. Alignments were generated using a Clustal W-based algorithm within the Vector NTI software package (Invitrogen). Positions of PCR primer pairs for ChIP amplification are also shown. H. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 (a component of the NuRD complex) and Jmjd1a. The PCR primer pair spanned the human Aldh1a1 octamer site. ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.
    Figure Legend Snippet: A. Aldefluor staining profile of A549 cells infected with a doxycycline-inducible lentiviral shRNA. MFI was 169.0 for scrambled and 91.2 for Oct1-specific shRNA. A549 cells were infected with control or Oct1-specific lentiviral particles (Santa Cruz), selected with puromycin, and subjected to analysis after 48 hr. B. Efficacy of the A549 knockdown as assessed by Western blotting using anti-Oct1 antibodies and an anti-GAPDH loading control. C. Oct1 Western blot of unsorted normal A549 cells cultured under normal conditions, and sorted Aldefluor HI and Aldefluor LO populations is shown. GAPDH is used as a loading control. D. The same sorted cells or unsorted cells were subjected to qRT-PCR to determine Oct1 mRNA levels. Levels are show relative to GAPDH. Error bars depict standard deviations. E. Oct1 was ectopically expressed in A549 cells using a retrovirus (pBabe-Oct1) or empty vector. The mixed population of cells was subjected to selection with puromycin, and ALDH activity determined. F. Western blot using anti-Oct1 antibodies of the same cells shown in (E). ß-actin is shown as a loading control. G. Alignment of the Aldh1a1 promoter regions in several example vertebrate species. The conserved perfect octamer sequence centered at approximately −55 bp relative to the transcription start site is highlighted. Alignments were generated using a Clustal W-based algorithm within the Vector NTI software package (Invitrogen). Positions of PCR primer pairs for ChIP amplification are also shown. H. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 (a component of the NuRD complex) and Jmjd1a. The PCR primer pair spanned the human Aldh1a1 octamer site. ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Techniques Used: Staining, Infection, shRNA, Control, Knockdown, Western Blot, Cell Culture, Quantitative RT-PCR, Plasmid Preparation, Selection, Activity Assay, Sequencing, Generated, Software, Amplification

    A. SP assay of luciferase-positive A549 cells carrying an inducible scrambled or Oct1-specific shRNA . Cells were treated with doxycycline for 4 days. Side population assays were conducted as described in the section. B. Western blot showing Oct1 protein levels in the two cell lines from (A) with and without 4-day culture in doxycycline. ß-actin is shown as a loading control. C. Quantification of the changed in SP percentage from three independent trials. Error bars denote standard deviations. D. Alignment of Abcg2 first intron regions in human and mouse. The perfect octamer sequence in both species is highlighted. Human Abcg2 has two annotated transcription start sites, so the element is located at both +14 kb and −12.4 kb relative to the transcription start sites. E. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 and Jmjd1a. The PCR primer pair spanned the human Oct1 binding site in Abcg2 . ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.
    Figure Legend Snippet: A. SP assay of luciferase-positive A549 cells carrying an inducible scrambled or Oct1-specific shRNA . Cells were treated with doxycycline for 4 days. Side population assays were conducted as described in the section. B. Western blot showing Oct1 protein levels in the two cell lines from (A) with and without 4-day culture in doxycycline. ß-actin is shown as a loading control. C. Quantification of the changed in SP percentage from three independent trials. Error bars denote standard deviations. D. Alignment of Abcg2 first intron regions in human and mouse. The perfect octamer sequence in both species is highlighted. Human Abcg2 has two annotated transcription start sites, so the element is located at both +14 kb and −12.4 kb relative to the transcription start sites. E. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 and Jmjd1a. The PCR primer pair spanned the human Oct1 binding site in Abcg2 . ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Techniques Used: Luciferase, shRNA, Western Blot, Control, Sequencing, Binding Assay

    A. Numbers of nude mice engrafted in the left flank with A549 cells expressing the same Oct1-specific shRNA as shown in . Scrambled shRNAs in the contralateral flank were used as controls. The fraction of mice successfully engrafted is shown in tabular format. For the Oct1-specific and control shRNA lines, calculation of TIC frequency is shown at bottom. B. Images of engrafted tumors from animals receiving 1×10 5 cells. Left flank: scrambled shRNA. Right flank: Oct1-specific shRNA. C. Similar to (A), except human Oct1 was over-expressed in luciferase-expressing A549 cells using retroviral gene transduction. D. 4 th inguinal mammary fat pads of nude mice were engrafted with the indicated number of luciferase-expressing MDA-MB-231 cells expressing scrambled or Oct1-specific shRNAs. The fraction of engrafted animals is shown in tabular format. E. Western blot showing efficacy of lentiviral knockdown. ß-actin is shown as a loading control. F. Images of engrafted tumors from animals receiving 2×10 5 cells. Ventral view is shown. Right side: scrambled shRNAs. Left side: Oct1-specific shRNAs. G. Similar to (D) except human Oct1 was overexpressed using retroviral gene transduction. H. Western blot showing degree of Oct1 overexpression in cells used in (G). ß-actin is shown as a loading control.
    Figure Legend Snippet: A. Numbers of nude mice engrafted in the left flank with A549 cells expressing the same Oct1-specific shRNA as shown in . Scrambled shRNAs in the contralateral flank were used as controls. The fraction of mice successfully engrafted is shown in tabular format. For the Oct1-specific and control shRNA lines, calculation of TIC frequency is shown at bottom. B. Images of engrafted tumors from animals receiving 1×10 5 cells. Left flank: scrambled shRNA. Right flank: Oct1-specific shRNA. C. Similar to (A), except human Oct1 was over-expressed in luciferase-expressing A549 cells using retroviral gene transduction. D. 4 th inguinal mammary fat pads of nude mice were engrafted with the indicated number of luciferase-expressing MDA-MB-231 cells expressing scrambled or Oct1-specific shRNAs. The fraction of engrafted animals is shown in tabular format. E. Western blot showing efficacy of lentiviral knockdown. ß-actin is shown as a loading control. F. Images of engrafted tumors from animals receiving 2×10 5 cells. Ventral view is shown. Right side: scrambled shRNAs. Left side: Oct1-specific shRNAs. G. Similar to (D) except human Oct1 was overexpressed using retroviral gene transduction. H. Western blot showing degree of Oct1 overexpression in cells used in (G). ß-actin is shown as a loading control.

    Techniques Used: Expressing, shRNA, Control, Luciferase, Retroviral, Transduction, Western Blot, Knockdown, Over Expression

    A. Peripheral blood leukocyte B and T cell profiles from primary and secondary animals transplanted with Oct1 deficient and WT littermate control fetal liver (FL). Cells were stained with anti-Thy1.2 and anti-B220 antibodies to reveal T and B cells. Control Rag1 −/− and WT animals are shown at the top for comparison. B. B and T cell repopulation is shown as a percentage of total cells for 5 independent experiments. C. Flow cytometry plots showing degree of T cell reconstitution in peripheral blood leukocytes in four Rag1 −/− recipient animals. For each animal, WT (top panels) or littermate Oct1 deficient (bottom panels) fetal livers were mixed with Thy1.1 + /Thy1.2 + WT bone marrow (BM) and injected retro-orbitally. Peripheral blood repopulation was analyzed at 10 and 15 weeks. D. The same peripheral blood analysis as in (C) was performed using Ly5.1/2 instead of Thy1.1/2 as a congenic marker. A single mouse for each condition is shown. E. Averages of 4 WT and 4 Oct1 deficient competitive repopulations as performed in (D) are shown. Peripheral blood was analyzed at 5 and 15 weeks. Error bars depict +/− standard deviation. F. Bone marrow from the mice in (D) were gated on LSK and analyzed for Ly5 expression. G. Averages of the 4 WT and 4 Oct1 deficient competitive repopulations at the level of LSK bone marrow precursors.
    Figure Legend Snippet: A. Peripheral blood leukocyte B and T cell profiles from primary and secondary animals transplanted with Oct1 deficient and WT littermate control fetal liver (FL). Cells were stained with anti-Thy1.2 and anti-B220 antibodies to reveal T and B cells. Control Rag1 −/− and WT animals are shown at the top for comparison. B. B and T cell repopulation is shown as a percentage of total cells for 5 independent experiments. C. Flow cytometry plots showing degree of T cell reconstitution in peripheral blood leukocytes in four Rag1 −/− recipient animals. For each animal, WT (top panels) or littermate Oct1 deficient (bottom panels) fetal livers were mixed with Thy1.1 + /Thy1.2 + WT bone marrow (BM) and injected retro-orbitally. Peripheral blood repopulation was analyzed at 10 and 15 weeks. D. The same peripheral blood analysis as in (C) was performed using Ly5.1/2 instead of Thy1.1/2 as a congenic marker. A single mouse for each condition is shown. E. Averages of 4 WT and 4 Oct1 deficient competitive repopulations as performed in (D) are shown. Peripheral blood was analyzed at 5 and 15 weeks. Error bars depict +/− standard deviation. F. Bone marrow from the mice in (D) were gated on LSK and analyzed for Ly5 expression. G. Averages of the 4 WT and 4 Oct1 deficient competitive repopulations at the level of LSK bone marrow precursors.

    Techniques Used: Control, Staining, Comparison, Flow Cytometry, Injection, Marker, Standard Deviation, Expressing



    Similar Products

    mixed rabbit anti oct1 antibodies  (Bethyl)
    93
    Bethyl mixed rabbit anti oct1 antibodies
    A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse <t>anti-Oct1</t> antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.
    Mixed Rabbit Anti Oct1 Antibodies, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mixed+rabbit+anti+oct1+antibodies/pmc03493455-226-6-10?v=Bethyl
    Average 93 stars, based on 1 article reviews
    mixed rabbit anti oct1 antibodies - by Bioz Stars, 2026-07
    93/100 stars
    		  Buy from Supplier

    Image Search Results


    A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. IF images of cross-sections from grossly uninvolved colon margins of a male familial adenomatous polyposis patient. Frozen sections were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and co-stained with TO-PRO. Crypts are shown in cross-section. White dashed circle highlights a crypt. Arrows indicate cells staining strongly for Oct1. B. IF images of colon crypt sections from a normal male individual. Sections were stained with DAPI, and anti-Oct1 and anti-ALDH1a1 antibodies. Merged images are shown at right. White dashed lines highlight crypts. Examples of cells co-staining with Oct1 and ALDH1a1 are highlighted with yellow arrows. An example cell staining with ALDH1a1 only is highlighted with an asterisk. Inset at lower right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. C. Frozen mouse colon tissue sections were stained with DAPI, and anti-Lrig1 and anti-Oct1 antibodies. IF images of longitudinal sections are shown. White dashed lines highlight the crypt. D. IF images of mouse small intestine sections. Sections were stained with DAPI and anti-Oct1 antibodies. Merged images are shown at right. White dashed lines highlight a crypt. Inset at upper right-hand corner is a digital magnification of the central portion of the image. Sections were formalin-fixed and paraffin-embedded. E. IF images of cross-sectional duodenum sections from a normal C57BL/6 mouse. Frozen sections were stained with DAPI and anti-Oct1 and anti-Lgr5 antibodies. Merged images are shown at right. Examples of co-staining cells are highlighted with yellow arrows. White dashed lines highlights crypts. F. Longitudinal sections.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Staining

    A. IF images are shown. Frozen malignant breast carcinoma sections (non-familial stage IIIA node-positive infiltrating ductal carcinoma, ER + PR + HER2 − ) were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and rabbit anti-ALDH1 antibodies (Abcam ab52492). Arrow indicates example double-positive cell. Asterisks show examples of an Oct1 HI cell with low ALDH1 and ALDH1 HI with low Oct1. Detail is shown in images below. B. Western blot depicting Oct1 levels in a panel of malignant epithelial cells isolated from pleural effusions (human breast carcinoma lung metastases). GAPDH is shown as a loading control. C. Examples of the CD24/44 profile from pleural effusions with highest and lowest Oct1 protein levels. D. Correlation of Oct1 protein and CD24 LO CD44 HI stem content in 15 individual patient samples (7 ER + PR + HER2 − , 5 ER − PR − HER2 − , 3 ER − PR − HER2 + ) collected from 14 different patients. One patient had tumor cells collected twice 22 months apart. Samples were placed into Oct1 LO and Oct1 HI categories based on Oct1 Western blot signal intensity relative to GAPDH staining. Cells were tested for percentage CD24 LO CD44 HI stem cell content and plotted. P -value was calculated using the two-tailed student T-test. E. The same analysis for Oct1 mRNA as measured by qRT-PCR.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. IF images are shown. Frozen malignant breast carcinoma sections (non-familial stage IIIA node-positive infiltrating ductal carcinoma, ER + PR + HER2 − ) were stained with mouse anti-Oct1 antibodies (Millipore MAB5434) and rabbit anti-ALDH1 antibodies (Abcam ab52492). Arrow indicates example double-positive cell. Asterisks show examples of an Oct1 HI cell with low ALDH1 and ALDH1 HI with low Oct1. Detail is shown in images below. B. Western blot depicting Oct1 levels in a panel of malignant epithelial cells isolated from pleural effusions (human breast carcinoma lung metastases). GAPDH is shown as a loading control. C. Examples of the CD24/44 profile from pleural effusions with highest and lowest Oct1 protein levels. D. Correlation of Oct1 protein and CD24 LO CD44 HI stem content in 15 individual patient samples (7 ER + PR + HER2 − , 5 ER − PR − HER2 − , 3 ER − PR − HER2 + ) collected from 14 different patients. One patient had tumor cells collected twice 22 months apart. Samples were placed into Oct1 LO and Oct1 HI categories based on Oct1 Western blot signal intensity relative to GAPDH staining. Cells were tested for percentage CD24 LO CD44 HI stem cell content and plotted. P -value was calculated using the two-tailed student T-test. E. The same analysis for Oct1 mRNA as measured by qRT-PCR.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Staining, Western Blot, Isolation, Control, Two Tailed Test, Quantitative RT-PCR

    A. Aldefluor staining profile of A549 cells infected with a doxycycline-inducible lentiviral shRNA. MFI was 169.0 for scrambled and 91.2 for Oct1-specific shRNA. A549 cells were infected with control or Oct1-specific lentiviral particles (Santa Cruz), selected with puromycin, and subjected to analysis after 48 hr. B. Efficacy of the A549 knockdown as assessed by Western blotting using anti-Oct1 antibodies and an anti-GAPDH loading control. C. Oct1 Western blot of unsorted normal A549 cells cultured under normal conditions, and sorted Aldefluor HI and Aldefluor LO populations is shown. GAPDH is used as a loading control. D. The same sorted cells or unsorted cells were subjected to qRT-PCR to determine Oct1 mRNA levels. Levels are show relative to GAPDH. Error bars depict standard deviations. E. Oct1 was ectopically expressed in A549 cells using a retrovirus (pBabe-Oct1) or empty vector. The mixed population of cells was subjected to selection with puromycin, and ALDH activity determined. F. Western blot using anti-Oct1 antibodies of the same cells shown in (E). ß-actin is shown as a loading control. G. Alignment of the Aldh1a1 promoter regions in several example vertebrate species. The conserved perfect octamer sequence centered at approximately −55 bp relative to the transcription start site is highlighted. Alignments were generated using a Clustal W-based algorithm within the Vector NTI software package (Invitrogen). Positions of PCR primer pairs for ChIP amplification are also shown. H. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 (a component of the NuRD complex) and Jmjd1a. The PCR primer pair spanned the human Aldh1a1 octamer site. ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. Aldefluor staining profile of A549 cells infected with a doxycycline-inducible lentiviral shRNA. MFI was 169.0 for scrambled and 91.2 for Oct1-specific shRNA. A549 cells were infected with control or Oct1-specific lentiviral particles (Santa Cruz), selected with puromycin, and subjected to analysis after 48 hr. B. Efficacy of the A549 knockdown as assessed by Western blotting using anti-Oct1 antibodies and an anti-GAPDH loading control. C. Oct1 Western blot of unsorted normal A549 cells cultured under normal conditions, and sorted Aldefluor HI and Aldefluor LO populations is shown. GAPDH is used as a loading control. D. The same sorted cells or unsorted cells were subjected to qRT-PCR to determine Oct1 mRNA levels. Levels are show relative to GAPDH. Error bars depict standard deviations. E. Oct1 was ectopically expressed in A549 cells using a retrovirus (pBabe-Oct1) or empty vector. The mixed population of cells was subjected to selection with puromycin, and ALDH activity determined. F. Western blot using anti-Oct1 antibodies of the same cells shown in (E). ß-actin is shown as a loading control. G. Alignment of the Aldh1a1 promoter regions in several example vertebrate species. The conserved perfect octamer sequence centered at approximately −55 bp relative to the transcription start site is highlighted. Alignments were generated using a Clustal W-based algorithm within the Vector NTI software package (Invitrogen). Positions of PCR primer pairs for ChIP amplification are also shown. H. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 (a component of the NuRD complex) and Jmjd1a. The PCR primer pair spanned the human Aldh1a1 octamer site. ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Staining, Infection, shRNA, Control, Knockdown, Western Blot, Cell Culture, Quantitative RT-PCR, Plasmid Preparation, Selection, Activity Assay, Sequencing, Generated, Software, Amplification

    A. SP assay of luciferase-positive A549 cells carrying an inducible scrambled or Oct1-specific shRNA . Cells were treated with doxycycline for 4 days. Side population assays were conducted as described in the section. B. Western blot showing Oct1 protein levels in the two cell lines from (A) with and without 4-day culture in doxycycline. ß-actin is shown as a loading control. C. Quantification of the changed in SP percentage from three independent trials. Error bars denote standard deviations. D. Alignment of Abcg2 first intron regions in human and mouse. The perfect octamer sequence in both species is highlighted. Human Abcg2 has two annotated transcription start sites, so the element is located at both +14 kb and −12.4 kb relative to the transcription start sites. E. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 and Jmjd1a. The PCR primer pair spanned the human Oct1 binding site in Abcg2 . ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. SP assay of luciferase-positive A549 cells carrying an inducible scrambled or Oct1-specific shRNA . Cells were treated with doxycycline for 4 days. Side population assays were conducted as described in the section. B. Western blot showing Oct1 protein levels in the two cell lines from (A) with and without 4-day culture in doxycycline. ß-actin is shown as a loading control. C. Quantification of the changed in SP percentage from three independent trials. Error bars denote standard deviations. D. Alignment of Abcg2 first intron regions in human and mouse. The perfect octamer sequence in both species is highlighted. Human Abcg2 has two annotated transcription start sites, so the element is located at both +14 kb and −12.4 kb relative to the transcription start sites. E. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 and Jmjd1a. The PCR primer pair spanned the human Oct1 binding site in Abcg2 . ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the section. Values are the average of four independent experiments. Error bars represent standard deviations.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Luciferase, shRNA, Western Blot, Control, Sequencing, Binding Assay

    A. Numbers of nude mice engrafted in the left flank with A549 cells expressing the same Oct1-specific shRNA as shown in . Scrambled shRNAs in the contralateral flank were used as controls. The fraction of mice successfully engrafted is shown in tabular format. For the Oct1-specific and control shRNA lines, calculation of TIC frequency is shown at bottom. B. Images of engrafted tumors from animals receiving 1×10 5 cells. Left flank: scrambled shRNA. Right flank: Oct1-specific shRNA. C. Similar to (A), except human Oct1 was over-expressed in luciferase-expressing A549 cells using retroviral gene transduction. D. 4 th inguinal mammary fat pads of nude mice were engrafted with the indicated number of luciferase-expressing MDA-MB-231 cells expressing scrambled or Oct1-specific shRNAs. The fraction of engrafted animals is shown in tabular format. E. Western blot showing efficacy of lentiviral knockdown. ß-actin is shown as a loading control. F. Images of engrafted tumors from animals receiving 2×10 5 cells. Ventral view is shown. Right side: scrambled shRNAs. Left side: Oct1-specific shRNAs. G. Similar to (D) except human Oct1 was overexpressed using retroviral gene transduction. H. Western blot showing degree of Oct1 overexpression in cells used in (G). ß-actin is shown as a loading control.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. Numbers of nude mice engrafted in the left flank with A549 cells expressing the same Oct1-specific shRNA as shown in . Scrambled shRNAs in the contralateral flank were used as controls. The fraction of mice successfully engrafted is shown in tabular format. For the Oct1-specific and control shRNA lines, calculation of TIC frequency is shown at bottom. B. Images of engrafted tumors from animals receiving 1×10 5 cells. Left flank: scrambled shRNA. Right flank: Oct1-specific shRNA. C. Similar to (A), except human Oct1 was over-expressed in luciferase-expressing A549 cells using retroviral gene transduction. D. 4 th inguinal mammary fat pads of nude mice were engrafted with the indicated number of luciferase-expressing MDA-MB-231 cells expressing scrambled or Oct1-specific shRNAs. The fraction of engrafted animals is shown in tabular format. E. Western blot showing efficacy of lentiviral knockdown. ß-actin is shown as a loading control. F. Images of engrafted tumors from animals receiving 2×10 5 cells. Ventral view is shown. Right side: scrambled shRNAs. Left side: Oct1-specific shRNAs. G. Similar to (D) except human Oct1 was overexpressed using retroviral gene transduction. H. Western blot showing degree of Oct1 overexpression in cells used in (G). ß-actin is shown as a loading control.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Expressing, shRNA, Control, Luciferase, Retroviral, Transduction, Western Blot, Knockdown, Over Expression

    A. Peripheral blood leukocyte B and T cell profiles from primary and secondary animals transplanted with Oct1 deficient and WT littermate control fetal liver (FL). Cells were stained with anti-Thy1.2 and anti-B220 antibodies to reveal T and B cells. Control Rag1 −/− and WT animals are shown at the top for comparison. B. B and T cell repopulation is shown as a percentage of total cells for 5 independent experiments. C. Flow cytometry plots showing degree of T cell reconstitution in peripheral blood leukocytes in four Rag1 −/− recipient animals. For each animal, WT (top panels) or littermate Oct1 deficient (bottom panels) fetal livers were mixed with Thy1.1 + /Thy1.2 + WT bone marrow (BM) and injected retro-orbitally. Peripheral blood repopulation was analyzed at 10 and 15 weeks. D. The same peripheral blood analysis as in (C) was performed using Ly5.1/2 instead of Thy1.1/2 as a congenic marker. A single mouse for each condition is shown. E. Averages of 4 WT and 4 Oct1 deficient competitive repopulations as performed in (D) are shown. Peripheral blood was analyzed at 5 and 15 weeks. Error bars depict +/− standard deviation. F. Bone marrow from the mice in (D) were gated on LSK and analyzed for Ly5 expression. G. Averages of the 4 WT and 4 Oct1 deficient competitive repopulations at the level of LSK bone marrow precursors.

    Journal: PLoS Genetics

    Article Title: Transcription Factor Oct1 Is a Somatic and Cancer Stem Cell Determinant

    doi: 10.1371/journal.pgen.1003048

    Figure Lengend Snippet: A. Peripheral blood leukocyte B and T cell profiles from primary and secondary animals transplanted with Oct1 deficient and WT littermate control fetal liver (FL). Cells were stained with anti-Thy1.2 and anti-B220 antibodies to reveal T and B cells. Control Rag1 −/− and WT animals are shown at the top for comparison. B. B and T cell repopulation is shown as a percentage of total cells for 5 independent experiments. C. Flow cytometry plots showing degree of T cell reconstitution in peripheral blood leukocytes in four Rag1 −/− recipient animals. For each animal, WT (top panels) or littermate Oct1 deficient (bottom panels) fetal livers were mixed with Thy1.1 + /Thy1.2 + WT bone marrow (BM) and injected retro-orbitally. Peripheral blood repopulation was analyzed at 10 and 15 weeks. D. The same peripheral blood analysis as in (C) was performed using Ly5.1/2 instead of Thy1.1/2 as a congenic marker. A single mouse for each condition is shown. E. Averages of 4 WT and 4 Oct1 deficient competitive repopulations as performed in (D) are shown. Peripheral blood was analyzed at 5 and 15 weeks. Error bars depict +/− standard deviation. F. Bone marrow from the mice in (D) were gated on LSK and analyzed for Ly5 expression. G. Averages of the 4 WT and 4 Oct1 deficient competitive repopulations at the level of LSK bone marrow precursors.

    Article Snippet: For panels 1D and 2A, two mixed rabbit anti-Oct1 antibodies (Bethyl, A301-716A, A301-717A) were used.

    Techniques: Control, Staining, Comparison, Flow Cytometry, Injection, Marker, Standard Deviation, Expressing