mhcc97-h c-met shrna cells Search Results


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    Thermo Fisher sirna
    c-Met regulates CD44s expression through AKT signaling. (A) <t>MHCC97-H</t> cells were treated with 1 μM of PHA665752 for 24 h. Immunoblot analysis of CD44v6 (~160 kDa), CD44s (~85 kDa), and B-actin. (B) MHCC97-H cells were treated with 25 μM DMSO, LY294002 (PI3K inhibitor) or PD98059 (MEK inhibitor) for 24 h, and immunoblot analysis was performed. (C) MHCC97-H cells were stably transfected with c-Met <t>shRNA.</t> Lysates were collected after 2 passages and CD44s, fibronectin, and E-cadherin expression was via immunoblotting. (D) Tumorsphere formation (40X magnification) assay of stably transfected MHCC97-H cells with c-Met shRNA compared to scrambled control. The data represent the mean ± SEM of triplicates, *p
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    Thermo Fisher lipofectamine rnaimax
    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post <t>transfection.</t> C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p
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    Promega fugene 6 transfection reagents
    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post <t>transfection.</t> C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p
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    Illumina Inc human gene chip
    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post <t>transfection.</t> C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p
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    Illumina Inc microarray analysis
    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post <t>transfection.</t> C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p
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    Illumina Inc transcriptome analysis
    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post <t>transfection.</t> C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p
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    c-Met regulates CD44s expression through AKT signaling. (A) MHCC97-H cells were treated with 1 μM of PHA665752 for 24 h. Immunoblot analysis of CD44v6 (~160 kDa), CD44s (~85 kDa), and B-actin. (B) MHCC97-H cells were treated with 25 μM DMSO, LY294002 (PI3K inhibitor) or PD98059 (MEK inhibitor) for 24 h, and immunoblot analysis was performed. (C) MHCC97-H cells were stably transfected with c-Met shRNA. Lysates were collected after 2 passages and CD44s, fibronectin, and E-cadherin expression was via immunoblotting. (D) Tumorsphere formation (40X magnification) assay of stably transfected MHCC97-H cells with c-Met shRNA compared to scrambled control. The data represent the mean ± SEM of triplicates, *p

    Journal: BMC Cancer

    Article Title: Induction of tumor initiation is dependent on CD44s in c-Met+ hepatocellular carcinoma

    doi: 10.1186/s12885-015-1166-4

    Figure Lengend Snippet: c-Met regulates CD44s expression through AKT signaling. (A) MHCC97-H cells were treated with 1 μM of PHA665752 for 24 h. Immunoblot analysis of CD44v6 (~160 kDa), CD44s (~85 kDa), and B-actin. (B) MHCC97-H cells were treated with 25 μM DMSO, LY294002 (PI3K inhibitor) or PD98059 (MEK inhibitor) for 24 h, and immunoblot analysis was performed. (C) MHCC97-H cells were stably transfected with c-Met shRNA. Lysates were collected after 2 passages and CD44s, fibronectin, and E-cadherin expression was via immunoblotting. (D) Tumorsphere formation (40X magnification) assay of stably transfected MHCC97-H cells with c-Met shRNA compared to scrambled control. The data represent the mean ± SEM of triplicates, *p

    Article Snippet: Briefly, 1X105 MHCC97-H cells were transfected with 25pM of c-Met siRNA or scrambled siRNA (Thermo Scientific, Dharmacon, Chicago, IL) followed by transfection of pBabe CD44s construct or pBabe EV and incubated for an additional 24 hours.

    Techniques: Expressing, Stable Transfection, Transfection, shRNA

    c-Met induces tumor-initiating characteristics through CD44s. (A) 5x10 4 MHCC97-H cells were plated onto 6-well low adherent culture plates in triplicates. Tumorspheres were collected by centrifugation after 2 weeks . For monolayer cultured cells, 5x10 4 MHCC97-H were plated on 10 cm plate and cultured for 2 weeks. Media was changed every 2–3 days. MHCC97-H cells were plated in low-adherent cell or monolayer cell culture dishes for two weeks and immunoblotting analysis was confirmed on tumorsphere lysates. The data are representative of three independent experiments. (B) 5x10 3 MHCC97-H scrambled, c-Met shRNA and CD44s shRNA cells were grown in 6-well low-adherent culture plates for two weeks and the numbers of tumorspheres were counted (40X magnification). The data are representative of two independent experiments and are shown as the mean ± SEM of triplicate plates. (C) CD44s recovers tumorsphere formation. MHCC97-H cells were transfected with c-Met siRNA (25pM) for 24 hrs followed by overexpression of CD44s or pBabe empty vector retrovirus for an additional 48 h (immunoblot) or two weeks (tumorsphere assay). (D) Immunoblot data are representative of two independent experiments. The data for the tumorsphere assay data are representative of two independent experiments and are shown as the mean ± SEM of triplicate wells (40X magnification).

    Journal: BMC Cancer

    Article Title: Induction of tumor initiation is dependent on CD44s in c-Met+ hepatocellular carcinoma

    doi: 10.1186/s12885-015-1166-4

    Figure Lengend Snippet: c-Met induces tumor-initiating characteristics through CD44s. (A) 5x10 4 MHCC97-H cells were plated onto 6-well low adherent culture plates in triplicates. Tumorspheres were collected by centrifugation after 2 weeks . For monolayer cultured cells, 5x10 4 MHCC97-H were plated on 10 cm plate and cultured for 2 weeks. Media was changed every 2–3 days. MHCC97-H cells were plated in low-adherent cell or monolayer cell culture dishes for two weeks and immunoblotting analysis was confirmed on tumorsphere lysates. The data are representative of three independent experiments. (B) 5x10 3 MHCC97-H scrambled, c-Met shRNA and CD44s shRNA cells were grown in 6-well low-adherent culture plates for two weeks and the numbers of tumorspheres were counted (40X magnification). The data are representative of two independent experiments and are shown as the mean ± SEM of triplicate plates. (C) CD44s recovers tumorsphere formation. MHCC97-H cells were transfected with c-Met siRNA (25pM) for 24 hrs followed by overexpression of CD44s or pBabe empty vector retrovirus for an additional 48 h (immunoblot) or two weeks (tumorsphere assay). (D) Immunoblot data are representative of two independent experiments. The data for the tumorsphere assay data are representative of two independent experiments and are shown as the mean ± SEM of triplicate wells (40X magnification).

    Article Snippet: Briefly, 1X105 MHCC97-H cells were transfected with 25pM of c-Met siRNA or scrambled siRNA (Thermo Scientific, Dharmacon, Chicago, IL) followed by transfection of pBabe CD44s construct or pBabe EV and incubated for an additional 24 hours.

    Techniques: Centrifugation, Cell Culture, shRNA, Transfection, Over Expression, Plasmid Preparation

    CD44s regulates tumor initiation in vivo . (A) Immunoblot of MHCC97-H scrambled and CD44s shRNA #1 before cells were injected into athymic nude mice. (B-D) Tumor initiation graph of MHCC97-H CD44s shRNA compared with the scrambled shRNA control. Bilateral subcutaneous injections of 1x10 2 , 1x10 3 , or 1x10 4 cells were inoculated into athymic nude mice, and the number of tumors formed and the percent tumor initiation were calculated (1x10 2 , N = 10; 1x10 3 , N = 8; or 1x10 4 , N = 6). Tumor volume was calculated at the end of the experiment and. Data represent the mean ± SD. Confirmation of down-regulation of CD44s and c-Met signaling was performed by immunoblotting.

    Journal: BMC Cancer

    Article Title: Induction of tumor initiation is dependent on CD44s in c-Met+ hepatocellular carcinoma

    doi: 10.1186/s12885-015-1166-4

    Figure Lengend Snippet: CD44s regulates tumor initiation in vivo . (A) Immunoblot of MHCC97-H scrambled and CD44s shRNA #1 before cells were injected into athymic nude mice. (B-D) Tumor initiation graph of MHCC97-H CD44s shRNA compared with the scrambled shRNA control. Bilateral subcutaneous injections of 1x10 2 , 1x10 3 , or 1x10 4 cells were inoculated into athymic nude mice, and the number of tumors formed and the percent tumor initiation were calculated (1x10 2 , N = 10; 1x10 3 , N = 8; or 1x10 4 , N = 6). Tumor volume was calculated at the end of the experiment and. Data represent the mean ± SD. Confirmation of down-regulation of CD44s and c-Met signaling was performed by immunoblotting.

    Article Snippet: Briefly, 1X105 MHCC97-H cells were transfected with 25pM of c-Met siRNA or scrambled siRNA (Thermo Scientific, Dharmacon, Chicago, IL) followed by transfection of pBabe CD44s construct or pBabe EV and incubated for an additional 24 hours.

    Techniques: In Vivo, shRNA, Injection, Mouse Assay

    siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post transfection. C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p

    Journal: PLoS ONE

    Article Title: The EGFR/ErbB3 Pathway Acts as a Compensatory Survival Mechanism upon c-Met Inhibition in Human c-Met+ Hepatocellular Carcinoma

    doi: 10.1371/journal.pone.0128159

    Figure Lengend Snippet: siRNA screening and microarray analysis of MHCC97-H liver cancer cell line stably transfected with c-Met shRNA reveals EGFR pathway as a putative survival pathway in HCC. A) c-Met shRNA was stably transfected into the MHCC97-H cell line, which has constitutive c-Met activity. After puromycin selection, immunoblot determined c-Met knockdown in a c-Met + HCC cell line suppresses downstream signaling (c-Met, Akt, and Erk1/2 phosphorylation) compared to MHCC97-H cells stably expressing a scrambled shRNA. B) An XTT assay was performed to confirm the eight targets from the siRNA screen that had the greatest effect on cell viability in MHCC97-H c-Met KD cells. 10 nM siRNA and 0.2 ul RNAiMAX were used to transfect MHCC97-H c-Met KD cells and cell viability was determined at 48 hours post transfection. C) Ingenuity pathway analysis was conducted to compare microarray gene expression between MHCC97-H c-Met knockdown (KD) cells and MHCC97-H cells stably expressing a scrambled shRNA. The top seven enriched pathways are shown. D) A heatmap of the subset of the EGFR pathway gene set that is differentially expressed by microarray (Illumina human gene chip). A statistically significant (p

    Article Snippet: 5×103 MHCC97-H c-Met shRNA cells were plated in 96-well plates and reverse transfected (cells were added to 10 nM siRNA and 0.2 μl RNAiMAX pre-added to wells) with individual siRNA using lipid-mediated transfection with Lipofectamine RNAiMAX (Life Technologies Corporation, Grand Island, NY).

    Techniques: Microarray, Stable Transfection, Transfection, shRNA, Activity Assay, Selection, Expressing, XTT Assay, Chromatin Immunoprecipitation