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Polysciences inc pei max
WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated <t>pmCherry-SAMHD1</t> K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using <t>PEI</t> Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.
Pei Max, supplied by Polysciences inc, 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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pei max - by Bioz Stars, 2021-09
86/100 stars

Images

1) Product Images from "Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation"

Article Title: Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0066201

WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.
Figure Legend Snippet: WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.

Techniques Used: Transfection, Incubation, Lysis, SDS Page

Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.
Figure Legend Snippet: Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.

Techniques Used: Mutagenesis, Live Cell Imaging, Plasmid Preparation, Transfection, Cell Culture, Imaging, Software

2) Product Images from "Abstracts from the 25th European Society for Animal Cell Technology Meeting: Cell Technologies for Innovative Therapies"

Article Title: Abstracts from the 25th European Society for Animal Cell Technology Meeting: Cell Technologies for Innovative Therapies

Journal: BMC Proceedings

doi: 10.1186/s12919-018-0097-x

PEIpro® requires less reagent and similar to lower DNA amount compared to other PEIs. Suspension HEK-293 and CHO cells were seeded at 1×10 6 cells/mL in serum free medium and transfected with PEIpro®, PEI “Max” and L-PEI 25 kDa (Polysciences, Warrington, PA) resuspended at 1 mg/ml. Luciferase expression was assayed 48 h after transfection using a conventional luciferase assay
Figure Legend Snippet: PEIpro® requires less reagent and similar to lower DNA amount compared to other PEIs. Suspension HEK-293 and CHO cells were seeded at 1×10 6 cells/mL in serum free medium and transfected with PEIpro®, PEI “Max” and L-PEI 25 kDa (Polysciences, Warrington, PA) resuspended at 1 mg/ml. Luciferase expression was assayed 48 h after transfection using a conventional luciferase assay

Techniques Used: Transfection, Luciferase, Expressing

3) Product Images from "Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation"

Article Title: Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0066201

WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.
Figure Legend Snippet: WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.

Techniques Used: Transfection, Incubation, Lysis, SDS Page

Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.
Figure Legend Snippet: Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.

Techniques Used: Mutagenesis, Live Cell Imaging, Plasmid Preparation, Transfection, Cell Culture, Imaging, Software

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Plasmid Preparation:

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Cell Culture:

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Live Cell Imaging:

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    Polysciences inc pei max
    WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated <t>pmCherry-SAMHD1</t> K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using <t>PEI</t> Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.
    Pei Max, supplied by Polysciences inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pei max/product/Polysciences inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pei max - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    86
    Polysciences inc polyethyleneimine pei max transfection reagent
    QR-110 Increases Wild-Type CEP290 mRNA and Protein Levels in c.2991+1655A > G Homozygous and Compound Heterozygous LCA Primary Fibroblasts (A–D) Quantification of (A) wild-type transcript (exon 26-27), (B) c.2991+1655A > G mutant transcript (exon X-27), and (C and D) protein in control and c.2991+1655A > G homozygous and compound heterozygous primary fibroblasts following treatment with QR-110. Control and LCA cells were treated with different concentrations of QR-110 using <t>PEI-mediated</t> <t>transfection</t> and harvested after 24 (RNA) or 72 hr (proteins). Mock: transfection reagent only; scrambled: different sequence control oligonucleotide with same length and chemistry. mRNA levels were determined by one-step isoform-specific RT-ddPCR. (C) Representative CEP290 immunoblot images from different LCA cell lines. (D) Protein signal was quantified (n = 3 biological replicates per cell line) by densitometry analysis, normalized by protein load (LC), measured by UV illumination of Stain-free gel, and expressed relative to control cell levels. Data are represented as mean ± SEM. *p
    Polyethyleneimine Pei Max Transfection Reagent, supplied by Polysciences inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyethyleneimine pei max transfection reagent/product/Polysciences inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polyethyleneimine pei max transfection reagent - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    Image Search Results


    WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.

    Journal: PLoS ONE

    Article Title: Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation

    doi: 10.1371/journal.pone.0066201

    Figure Lengend Snippet: WT SAMHD1 relocates cytoplasm-localized mutants of SAMHD1K11A to the nucleus and facilitates their Vpx-induced degradation. (A)–(C) Wild-type SAMHD1-HA partially translocated pmCherry-SAMHD1 K11A to the nucleus. pmCherry-SAMHD1-HA WT and/or K11A with pEYFP-Nuc were co-transfected into HEK293T cells using PEI Max. Live cells were imaged at 24 h post-transfection. (D) Interaction between wild-type SAMHD1 and K11A. pSAMHD1 WT-Flag was co-transfected with pmCherry-SAMHD1-HA WT or K11A. Transfected cells were harvested after 48 h and incubated in lysis buffer for 30 min. Cell lysates were added to an anti-HA affinity matrix (Roche). Eluted protein samples were detected by immunoblotting with anti-HA and anti-Flag antibody to detect mCherry-SAMHD1 and SAMHD1-Flag, respectively. (E) Immunoblotting demonstrates that pmCherry-SAMHD1 K11A became sensitive to Vpx-mediated degradation when co-expressed with SAMHD1 WT. HEK293T cells were co-transfected with HA-Vpx and pmCherrySAMHD1 K11A and/or pSAMHD1-Flag wild-type. Cell extracts were harvested 48 h later and analyzed by SDS-PAGE, followed by immunoblotting to detect SAMHD1-HA and HA-Vpx. β-actin was used as the loading control.

    Article Snippet: Live cell imaging Plasmid (pEYFP-Nuc [a gift of Dr. T. Inoue], 0.25 µg) and pmCherry-SAMHD1 -HA (2 µg) were transfected into HEK293T cells using PEI Max (Polysciences) according to the manufacturer's protocol.

    Techniques: Transfection, Incubation, Lysis, SDS Page

    Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.

    Journal: PLoS ONE

    Article Title: Identification of Critical Regions in Human SAMHD1 Required for Nuclear Localization and Vpx-Mediated Degradation

    doi: 10.1371/journal.pone.0066201

    Figure Lengend Snippet: Cellular localization of mCherry-tagged SAMHD1 and SAMHD1 mutant proteins using live cell imaging. (A)–(F) Plasmid pEYFP-Nuc (Clontech) and pmCherry-SAMHD1 WT or indicated variants were co-transfected into HEK293T cells using PEI Max (Polysciences). For live cell imaging, HEK293T cells were transfected in 6-well coverslip glass-bottomed cell culture dishes (InVitro Scientific) when the cells were ∼80% confluent, and then visualized after 24 h using a Zeiss LSM510-Meta confocal imaging system equipped with four argon lasers (458-, 477-, 488-, and 514-nm lines), two HeNe lasers (542 and 633 nm), and one diode laser (405 nm). All images were acquired from a 100X objective, and image analysis and manipulation were performed using Zen 2009 software.

    Article Snippet: Live cell imaging Plasmid (pEYFP-Nuc [a gift of Dr. T. Inoue], 0.25 µg) and pmCherry-SAMHD1 -HA (2 µg) were transfected into HEK293T cells using PEI Max (Polysciences) according to the manufacturer's protocol.

    Techniques: Mutagenesis, Live Cell Imaging, Plasmid Preparation, Transfection, Cell Culture, Imaging, Software

    PEIpro® requires less reagent and similar to lower DNA amount compared to other PEIs. Suspension HEK-293 and CHO cells were seeded at 1×10 6 cells/mL in serum free medium and transfected with PEIpro®, PEI “Max” and L-PEI 25 kDa (Polysciences, Warrington, PA) resuspended at 1 mg/ml. Luciferase expression was assayed 48 h after transfection using a conventional luciferase assay

    Journal: BMC Proceedings

    Article Title: Abstracts from the 25th European Society for Animal Cell Technology Meeting: Cell Technologies for Innovative Therapies

    doi: 10.1186/s12919-018-0097-x

    Figure Lengend Snippet: PEIpro® requires less reagent and similar to lower DNA amount compared to other PEIs. Suspension HEK-293 and CHO cells were seeded at 1×10 6 cells/mL in serum free medium and transfected with PEIpro®, PEI “Max” and L-PEI 25 kDa (Polysciences, Warrington, PA) resuspended at 1 mg/ml. Luciferase expression was assayed 48 h after transfection using a conventional luciferase assay

    Article Snippet: PEI max (Polysciences, Inc.) and BalanCD Transfectory CHO (Irvine Scientific) were used for the transfection.

    Techniques: Transfection, Luciferase, Expressing

    QR-110 Increases Wild-Type CEP290 mRNA and Protein Levels in c.2991+1655A > G Homozygous and Compound Heterozygous LCA Primary Fibroblasts (A–D) Quantification of (A) wild-type transcript (exon 26-27), (B) c.2991+1655A > G mutant transcript (exon X-27), and (C and D) protein in control and c.2991+1655A > G homozygous and compound heterozygous primary fibroblasts following treatment with QR-110. Control and LCA cells were treated with different concentrations of QR-110 using PEI-mediated transfection and harvested after 24 (RNA) or 72 hr (proteins). Mock: transfection reagent only; scrambled: different sequence control oligonucleotide with same length and chemistry. mRNA levels were determined by one-step isoform-specific RT-ddPCR. (C) Representative CEP290 immunoblot images from different LCA cell lines. (D) Protein signal was quantified (n = 3 biological replicates per cell line) by densitometry analysis, normalized by protein load (LC), measured by UV illumination of Stain-free gel, and expressed relative to control cell levels. Data are represented as mean ± SEM. *p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Splice-Modulating Oligonucleotide QR-110 Restores CEP290 mRNA and Function in Human c.2991+1655A > G LCA10 Models

    doi: 10.1016/j.omtn.2018.07.010

    Figure Lengend Snippet: QR-110 Increases Wild-Type CEP290 mRNA and Protein Levels in c.2991+1655A > G Homozygous and Compound Heterozygous LCA Primary Fibroblasts (A–D) Quantification of (A) wild-type transcript (exon 26-27), (B) c.2991+1655A > G mutant transcript (exon X-27), and (C and D) protein in control and c.2991+1655A > G homozygous and compound heterozygous primary fibroblasts following treatment with QR-110. Control and LCA cells were treated with different concentrations of QR-110 using PEI-mediated transfection and harvested after 24 (RNA) or 72 hr (proteins). Mock: transfection reagent only; scrambled: different sequence control oligonucleotide with same length and chemistry. mRNA levels were determined by one-step isoform-specific RT-ddPCR. (C) Representative CEP290 immunoblot images from different LCA cell lines. (D) Protein signal was quantified (n = 3 biological replicates per cell line) by densitometry analysis, normalized by protein load (LC), measured by UV illumination of Stain-free gel, and expressed relative to control cell levels. Data are represented as mean ± SEM. *p

    Article Snippet: Fibroblasts were transfected with oligonucleotides (see ) using polyethyleneimine (PEI) Max transfection reagent (Polysciences, Germany) with an oligo-to-PEI ratio of 1:4 and incubated for 24 hr (RNA analysis) or 3 days (protein analysis) at 37°C.

    Techniques: Mutagenesis, Transfection, Sequencing, Staining