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pxp2 backbone  (ATCC)


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    ATCC pxp2 backbone
    PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the <t>pXP2</t> luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.
    Pxp2 Backbone, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pxp2 backbone/product/ATCC
    Average 90 stars, based on 13 article reviews
    pxp2 backbone - by Bioz Stars, 2026-02
    90/100 stars

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    1) Product Images from "Macrophage development from HSCs requires PU.1-coordinated microRNA expression"

    Article Title: Macrophage development from HSCs requires PU.1-coordinated microRNA expression

    Journal: Blood

    doi: 10.1182/blood-2011-02-335141

    PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the pXP2 luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.
    Figure Legend Snippet: PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the pXP2 luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.

    Techniques Used: In Vivo, Expressing, Quantitative RT-PCR, Purification, Binding Assay, Luciferase, Plasmid Preparation, Mutagenesis, Construct, Activity Assay, Transfection, Fluorescence, shRNA, Stable Transfection, Western Blot



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    pxp2 backbone  (ATCC)
    90
    ATCC pxp2 backbone
    PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the <t>pXP2</t> luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.
    Pxp2 Backbone, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pxp2 backbone/product/ATCC
    Average 90 stars, based on 1 article reviews
    pxp2 backbone - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

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    PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the pXP2 luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.

    Journal: Blood

    Article Title: Macrophage development from HSCs requires PU.1-coordinated microRNA expression

    doi: 10.1182/blood-2011-02-335141

    Figure Lengend Snippet: PU.1 regulates miR-146a in vivo. (A) MiR-146a expression in WT hematopoietic precursors and mature myeloid cells. miR-146a qRT-PCR was performed from FACS-purified HSCs (LSKs), common myeloid progenitors (CMPs; Lin−Sca-1−c-Kit+CD34+FcγRII/IIIlow), splenic granulocytes (Gra; Gr1+CD11b+), and splenic macrophages (Mac; Gr1−CD11b+). Data were compared with U6 RNA ± SD miR-146a and PU.1 mRNA expression in the indicated FACS-purified hematopoietic populations from WT and PU.1 KD animals. (B) MiR-146a was analyzed compared with U6 RNA, and PU.1 expression was determined relative to β-actin. **P < .001 by Student t test. (C) A 375-bp fragment containing the PU.1-binding site at 10 kb upstream of the genomic miR-146 locus was inserted in the pXP2 luciferase plasmid pXP2-(-)10kb. Mutation of the PU.1 motif (ctgaggaagt to ctgaccaagt) in the pXP2-(-)10kb construct, as indicated by the crossed-out PU.1 symbol in the pictogram, led to a marked reduction in reporter activity in transiently transfected myeloid RAW264.7 cells. Relative expression was determined 48 hours after transfection compared with pXP2 empty basal fluorescence activity (n = 3). **P < .001 by Student t test. (D) RAW264.7 cells carrying a stable PU.1 KD shRNA showed significantly lower miR146 promoter activity than plain RAW264.7 cells when transiently transfected with the WT pXP2-(-)10kb construct. Reporter activity of plain RAW264.7 cells was set to 100% (n = 3). **P < .001 by Student t test. (E) PU.1 expression in untreated and stably PU.1-shRNA–transfected RAW264.7 cells in Western blot normalized to levels of tubulin. Error bars indicate SD.

    Article Snippet: A 375-bp genomic fragment located 10 kb upstream of miR-146a was PCR amplified (primer forward: 5′-ctcgaggagccggaatagaaggttcc-3′; primer reverse: 5′-aagcttaatgttaaattgaggtttttggtcg-3′) and subcloned into the pXP2 backbone (American Type Culture Collection no. 37577) via Xho I/ Hin dIII.

    Techniques: In Vivo, Expressing, Quantitative RT-PCR, Purification, Binding Assay, Luciferase, Plasmid Preparation, Mutagenesis, Construct, Activity Assay, Transfection, Fluorescence, shRNA, Stable Transfection, Western Blot