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pcmv sport6 cd63 phluorin  (Addgene inc)


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    Addgene inc pcmv sport6 cd63 phluorin
    Pcmv Sport6 Cd63 Phluorin, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcmv+sport6/pCMV-Sport6-CD63-pHluorin+(Plasmid+%23130901)/bio_rxiv__64898__2026__01__21__700874-48-7-8
    Average 93 stars, based on 23 article reviews
    pcmv sport6 cd63 phluorin - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Over Expression:

    Article Title: Myc manipulates the miRNA content and biologic functions of small cell lung cancer cell-derived small extracellular vesicles.
    Article Snippet: Background MYC genes are amplified/overexpressed in 20% of SCLCs, showing that Myc and Myc-dependent cellular mechanisms are strong candidates as therapeutic targets in SCLC.. Small extracellular vesicles support the carcinogenesis process by acting as messengers delivering nucleic acids and proteins—moreover, no reports associate Myc and the functional effect of small extracellular vesicles in small cell lung cancer.. Methods and results After the effects of small extracellular vesicles (sEVs) obtained from H82 and H209 cells on HUVEC and MRC-5 cells were observed, the Myc-dependent effect of the sEVs on oncogenic potentials was further evaluated by manipulating Myc expression via lentiviral vectors in H82 and H209 cells.

    Article Title: The effects of MYC on exosomes derived from cancer cells in the context of breast cancer.
    Article Snippet: Breast cancer is one of the cancer subtypes which account for most cancerrelated deaths in women each year globally (MassóVallés et al., 2022).. Breast cancer has multiple histological or molecular markers, and patients with breast cancer are classified clinically according to cancer cells' distinctive molecular features including expression Received: 9 December 2022 | Revised: 6 March 2023 | Accepted: 4 April 2023 DOI: 10.1111/cbdd.14245

    Transfection:

    Article Title: Myc manipulates the miRNA content and biologic functions of small cell lung cancer cell-derived small extracellular vesicles.
    Article Snippet: Background MYC genes are amplified/overexpressed in 20% of SCLCs, showing that Myc and Myc-dependent cellular mechanisms are strong candidates as therapeutic targets in SCLC.. Small extracellular vesicles support the carcinogenesis process by acting as messengers delivering nucleic acids and proteins—moreover, no reports associate Myc and the functional effect of small extracellular vesicles in small cell lung cancer.. Methods and results After the effects of small extracellular vesicles (sEVs) obtained from H82 and H209 cells on HUVEC and MRC-5 cells were observed, the Myc-dependent effect of the sEVs on oncogenic potentials was further evaluated by manipulating Myc expression via lentiviral vectors in H82 and H209 cells.

    Article Title: Real-time imaging of multivesicular body-plasma membrane fusion to quantify exosome release from single cells.
    Article Snippet: .. c CRITICAL The following TSPAN reporters are available from Addgene: ● CD63-pHluorin in pCMV-SPORT6 (Addgene ID 130901) ● CD81-pHluorin in pCMV-SPORT6 (Addgene ID 130903) ● CD9-pHluorin in pCMV-SPORT6 (Addgene ID 130905) ● CD63-pHuji in pCMV-SPORT6 (Addgene ID 130902) ● CD81-pHuji in pCMV-SPORT6 (Addgene ID 130904) ● CD9-pHuji in pCMV-SPORT6 (Addgene ID 130906) c CRITICAL TSPAN reporters in the pLenti6.3/TO/V5-DEST vector (for lentiviral transfection) are available upon request. .. Cell culture and transfection reagents ● DMEM + GlutaMAX-I (Gibco, cat. no. 61965-026) ● DMEM without phenol red (Gibco, cat. no. 31053-028) ● 10% (vol/vol) Fetal bovine serum (GE Healthcare Hyclone, cat. no. SV30160.03) ● Penicillin-streptomycin-glutamine (Gibco, cat. no. 10378-016) ● Trypsin-EDTA (Gibco, cat. no. 15400-054) ● Opti-MEM (Gibco, cat. no. 31985-070) ● Lipofectamine 2000 (Thermo Fisher, cat. no. 11668027) ● Poly-L-lysine 0.1 % (wt/vol) in H2O (Sigma-Aldrich, cat. no. P8920)

    Multiple Displacement Amplification:

    Article Title: The effects of MYC on exosomes derived from cancer cells in the context of breast cancer.
    Article Snippet: Breast cancer is one of the cancer subtypes which account for most cancerrelated deaths in women each year globally (MassóVallés et al., 2022).. Breast cancer has multiple histological or molecular markers, and patients with breast cancer are classified clinically according to cancer cells' distinctive molecular features including expression Received: 9 December 2022 | Revised: 6 March 2023 | Accepted: 4 April 2023 DOI: 10.1111/cbdd.14245

    Infection:

    Article Title: The effects of MYC on exosomes derived from cancer cells in the context of breast cancer.
    Article Snippet: Breast cancer is one of the cancer subtypes which account for most cancerrelated deaths in women each year globally (MassóVallés et al., 2022).. Breast cancer has multiple histological or molecular markers, and patients with breast cancer are classified clinically according to cancer cells' distinctive molecular features including expression Received: 9 December 2022 | Revised: 6 March 2023 | Accepted: 4 April 2023 DOI: 10.1111/cbdd.14245

    Plasmid Preparation:

    Article Title: Real-time imaging of multivesicular body-plasma membrane fusion to quantify exosome release from single cells.
    Article Snippet: .. c CRITICAL The following TSPAN reporters are available from Addgene: ● CD63-pHluorin in pCMV-SPORT6 (Addgene ID 130901) ● CD81-pHluorin in pCMV-SPORT6 (Addgene ID 130903) ● CD9-pHluorin in pCMV-SPORT6 (Addgene ID 130905) ● CD63-pHuji in pCMV-SPORT6 (Addgene ID 130902) ● CD81-pHuji in pCMV-SPORT6 (Addgene ID 130904) ● CD9-pHuji in pCMV-SPORT6 (Addgene ID 130906) c CRITICAL TSPAN reporters in the pLenti6.3/TO/V5-DEST vector (for lentiviral transfection) are available upon request. .. Cell culture and transfection reagents ● DMEM + GlutaMAX-I (Gibco, cat. no. 61965-026) ● DMEM without phenol red (Gibco, cat. no. 31053-028) ● 10% (vol/vol) Fetal bovine serum (GE Healthcare Hyclone, cat. no. SV30160.03) ● Penicillin-streptomycin-glutamine (Gibco, cat. no. 10378-016) ● Trypsin-EDTA (Gibco, cat. no. 15400-054) ● Opti-MEM (Gibco, cat. no. 31985-070) ● Lipofectamine 2000 (Thermo Fisher, cat. no. 11668027) ● Poly-L-lysine 0.1 % (wt/vol) in H2O (Sigma-Aldrich, cat. no. P8920)

    Article Title:
    Article Snippet: ATF4 5’UTR uORF1 and uORF2 mutations were introduced by PCR-based site-directed mutagenesis. pCMV-SPORT6 containing MGC Human MYC cDNA (CloneId:2985844) glycerol stock was obtained from Dharmacon (MHS6278). .. Fulllength MYC and GAPDH were amplified from cDNA and cloned into pCMV-SPORT6 using SalI and NotI restriction sites. pHR-EF1a-dCas9-HA-BFP-KRAB-NLS, a gift from the Jacob Corn Lab (Addgene plasmid #102244), was used to introduce catalytically dead Cas9 into HEK293T cell lines. ..

    Amplification:

    Article Title:
    Article Snippet: ATF4 5’UTR uORF1 and uORF2 mutations were introduced by PCR-based site-directed mutagenesis. pCMV-SPORT6 containing MGC Human MYC cDNA (CloneId:2985844) glycerol stock was obtained from Dharmacon (MHS6278). .. Fulllength MYC and GAPDH were amplified from cDNA and cloned into pCMV-SPORT6 using SalI and NotI restriction sites. pHR-EF1a-dCas9-HA-BFP-KRAB-NLS, a gift from the Jacob Corn Lab (Addgene plasmid #102244), was used to introduce catalytically dead Cas9 into HEK293T cell lines. ..

    Clone Assay:

    Article Title:
    Article Snippet: ATF4 5’UTR uORF1 and uORF2 mutations were introduced by PCR-based site-directed mutagenesis. pCMV-SPORT6 containing MGC Human MYC cDNA (CloneId:2985844) glycerol stock was obtained from Dharmacon (MHS6278). .. Fulllength MYC and GAPDH were amplified from cDNA and cloned into pCMV-SPORT6 using SalI and NotI restriction sites. pHR-EF1a-dCas9-HA-BFP-KRAB-NLS, a gift from the Jacob Corn Lab (Addgene plasmid #102244), was used to introduce catalytically dead Cas9 into HEK293T cell lines. ..

    Introduce:

    Article Title:
    Article Snippet: ATF4 5’UTR uORF1 and uORF2 mutations were introduced by PCR-based site-directed mutagenesis. pCMV-SPORT6 containing MGC Human MYC cDNA (CloneId:2985844) glycerol stock was obtained from Dharmacon (MHS6278). .. Fulllength MYC and GAPDH were amplified from cDNA and cloned into pCMV-SPORT6 using SalI and NotI restriction sites. pHR-EF1a-dCas9-HA-BFP-KRAB-NLS, a gift from the Jacob Corn Lab (Addgene plasmid #102244), was used to introduce catalytically dead Cas9 into HEK293T cell lines. ..



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    (A) Normalized fusion event frequencies under pharmacological and genetic modulation of PLD activity. A549 cells were transiently transfected <t>with</t> <t>CD63-pHmScarlet</t> and GFP-PASS, then treated with DMSO (control), the PLD1 inhibitor VU00155069 (PLD1i), or the dual PLD1/PLD2 inhibitor FIPI. Separately, cells were transfected with control siRNA, PLD1 siRNA, or a combination of PLD1 and PLD2 siRNAs. Fusion frequency was calculated as the number of fusion events per 100-second movie, then normalized to the control condition within each experimental day. Fusion frequencies were significantly reduced under PLD1i (*** p < 0.001), FIPI (** p < 0.01), PLD1 knockdown (* p < 0.05), and PLD1/2 knockdown (** p < 0.01) compared to their respective controls. (B) Quantification of membrane-proximal CD63 vesicle density at the plasma membrane. A549 cells co-expressing CD63-pHmScarlet and GFP-PASS were treated with 50 mM NH 4 Cl and imaged using TIRFM to enhance visualization. Membrane-proximal vesicles were defined as CD63-positive puncta visible in the TIRF field and detected using a custom MATLAB script employing bandpass filtering and intensity thresholding. Vesicle density was significantly reduced in PLD1i ( p = 0.014) and PLD1/2i ( p = 0.0299) conditions compared to DMSO. Each data point represents the vesicle density per cell area across three independent experiments. Cell counts: DMSO, n = 37; PLD1i, n = 39; PLD1/2i, n = 36. (C) Quantification of membrane-proximal lysosome density. A549 cells expressing CD63-pHluorin were stained with Magic Red and imaged by TIRFM. Cells were selected based on CD63-pHluorin expression. Membrane-proximal lysosomes were defined as Magic Red-positive puncta and detected using the same MATLAB-based analysis as in (B). Lysosome density was significantly increased under PLD1i ( p = 0.0028) but not significantly altered under PLD1/2i ( p = 0.2897) compared to DMSO. Each data point reflects the mean lysosome density per cell from three independent experiments ( n = 47 for all conditions). Statistical comparisons were performed using one-way ANOVA .
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    (A) Normalized fusion event frequencies under pharmacological and genetic modulation of PLD activity. A549 cells were transiently transfected <t>with</t> <t>CD63-pHmScarlet</t> and GFP-PASS, then treated with DMSO (control), the PLD1 inhibitor VU00155069 (PLD1i), or the dual PLD1/PLD2 inhibitor FIPI. Separately, cells were transfected with control siRNA, PLD1 siRNA, or a combination of PLD1 and PLD2 siRNAs. Fusion frequency was calculated as the number of fusion events per 100-second movie, then normalized to the control condition within each experimental day. Fusion frequencies were significantly reduced under PLD1i (*** p < 0.001), FIPI (** p < 0.01), PLD1 knockdown (* p < 0.05), and PLD1/2 knockdown (** p < 0.01) compared to their respective controls. (B) Quantification of membrane-proximal CD63 vesicle density at the plasma membrane. A549 cells co-expressing CD63-pHmScarlet and GFP-PASS were treated with 50 mM NH 4 Cl and imaged using TIRFM to enhance visualization. Membrane-proximal vesicles were defined as CD63-positive puncta visible in the TIRF field and detected using a custom MATLAB script employing bandpass filtering and intensity thresholding. Vesicle density was significantly reduced in PLD1i ( p = 0.014) and PLD1/2i ( p = 0.0299) conditions compared to DMSO. Each data point represents the vesicle density per cell area across three independent experiments. Cell counts: DMSO, n = 37; PLD1i, n = 39; PLD1/2i, n = 36. (C) Quantification of membrane-proximal lysosome density. A549 cells expressing CD63-pHluorin were stained with Magic Red and imaged by TIRFM. Cells were selected based on CD63-pHluorin expression. Membrane-proximal lysosomes were defined as Magic Red-positive puncta and detected using the same MATLAB-based analysis as in (B). Lysosome density was significantly increased under PLD1i ( p = 0.0028) but not significantly altered under PLD1/2i ( p = 0.2897) compared to DMSO. Each data point reflects the mean lysosome density per cell from three independent experiments ( n = 47 for all conditions). Statistical comparisons were performed using one-way ANOVA .
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    Image Search Results


    (A) Normalized fusion event frequencies under pharmacological and genetic modulation of PLD activity. A549 cells were transiently transfected with CD63-pHmScarlet and GFP-PASS, then treated with DMSO (control), the PLD1 inhibitor VU00155069 (PLD1i), or the dual PLD1/PLD2 inhibitor FIPI. Separately, cells were transfected with control siRNA, PLD1 siRNA, or a combination of PLD1 and PLD2 siRNAs. Fusion frequency was calculated as the number of fusion events per 100-second movie, then normalized to the control condition within each experimental day. Fusion frequencies were significantly reduced under PLD1i (*** p < 0.001), FIPI (** p < 0.01), PLD1 knockdown (* p < 0.05), and PLD1/2 knockdown (** p < 0.01) compared to their respective controls. (B) Quantification of membrane-proximal CD63 vesicle density at the plasma membrane. A549 cells co-expressing CD63-pHmScarlet and GFP-PASS were treated with 50 mM NH 4 Cl and imaged using TIRFM to enhance visualization. Membrane-proximal vesicles were defined as CD63-positive puncta visible in the TIRF field and detected using a custom MATLAB script employing bandpass filtering and intensity thresholding. Vesicle density was significantly reduced in PLD1i ( p = 0.014) and PLD1/2i ( p = 0.0299) conditions compared to DMSO. Each data point represents the vesicle density per cell area across three independent experiments. Cell counts: DMSO, n = 37; PLD1i, n = 39; PLD1/2i, n = 36. (C) Quantification of membrane-proximal lysosome density. A549 cells expressing CD63-pHluorin were stained with Magic Red and imaged by TIRFM. Cells were selected based on CD63-pHluorin expression. Membrane-proximal lysosomes were defined as Magic Red-positive puncta and detected using the same MATLAB-based analysis as in (B). Lysosome density was significantly increased under PLD1i ( p = 0.0028) but not significantly altered under PLD1/2i ( p = 0.2897) compared to DMSO. Each data point reflects the mean lysosome density per cell from three independent experiments ( n = 47 for all conditions). Statistical comparisons were performed using one-way ANOVA .

    Journal: bioRxiv

    Article Title: Phospholipase D1 and Phosphatidic Acid are required for MVE Fusion and Exosome Secretion

    doi: 10.64898/2026.01.21.700874

    Figure Lengend Snippet: (A) Normalized fusion event frequencies under pharmacological and genetic modulation of PLD activity. A549 cells were transiently transfected with CD63-pHmScarlet and GFP-PASS, then treated with DMSO (control), the PLD1 inhibitor VU00155069 (PLD1i), or the dual PLD1/PLD2 inhibitor FIPI. Separately, cells were transfected with control siRNA, PLD1 siRNA, or a combination of PLD1 and PLD2 siRNAs. Fusion frequency was calculated as the number of fusion events per 100-second movie, then normalized to the control condition within each experimental day. Fusion frequencies were significantly reduced under PLD1i (*** p < 0.001), FIPI (** p < 0.01), PLD1 knockdown (* p < 0.05), and PLD1/2 knockdown (** p < 0.01) compared to their respective controls. (B) Quantification of membrane-proximal CD63 vesicle density at the plasma membrane. A549 cells co-expressing CD63-pHmScarlet and GFP-PASS were treated with 50 mM NH 4 Cl and imaged using TIRFM to enhance visualization. Membrane-proximal vesicles were defined as CD63-positive puncta visible in the TIRF field and detected using a custom MATLAB script employing bandpass filtering and intensity thresholding. Vesicle density was significantly reduced in PLD1i ( p = 0.014) and PLD1/2i ( p = 0.0299) conditions compared to DMSO. Each data point represents the vesicle density per cell area across three independent experiments. Cell counts: DMSO, n = 37; PLD1i, n = 39; PLD1/2i, n = 36. (C) Quantification of membrane-proximal lysosome density. A549 cells expressing CD63-pHluorin were stained with Magic Red and imaged by TIRFM. Cells were selected based on CD63-pHluorin expression. Membrane-proximal lysosomes were defined as Magic Red-positive puncta and detected using the same MATLAB-based analysis as in (B). Lysosome density was significantly increased under PLD1i ( p = 0.0028) but not significantly altered under PLD1/2i ( p = 0.2897) compared to DMSO. Each data point reflects the mean lysosome density per cell from three independent experiments ( n = 47 for all conditions). Statistical comparisons were performed using one-way ANOVA .

    Article Snippet: The CD63-pHmScarlet construct was generated by modifying pCMV-Sport6-CD63-pHluorin (Addgene plasmid #130901, , replacing the pHluorin tag with pHmScarlet.

    Techniques: Activity Assay, Transfection, Control, Knockdown, Membrane, Clinical Proteomics, Expressing, Staining