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ATCC egfr wild type luad cell lines

Attenuation of YAP1 expression impairs cell proliferation and increases sensitivity to anti-cancer agents in <t>LUAD.</t> A, Western blotting confirmed YAP1-knockdown in <t>H1299</t> and <t>H1437</t> cells and YAP1-overexpression in <t>A549</t> cells. B, Proliferation assay: YAP1-deficient cells (H1299 YAP1-sh and H1437 YAP1-sh) showed lower proliferative ratios than did control cells, whereas YAP1-overexpressing cells (A549 YAP1-LV) proliferated more rapidly. C, Cells were treated with CDDP, and cell viability was recorded at 72 hours after treatment. H1299 and H1437 YAP1-sh cells showed greater sensitivity to CDDP, and YAP1-LV cells showed higher resistance, than did their control counterparts. D, YAP1-deficient cells treated with CDDP (2 μM or 5 μM for 72 hours) showed greater percentages of cells undergoing apoptosis than did control cells; whereas YAP1-LV cells showed substantially less apoptosis. E, Both YAP1-sh cell lines showed significantly suppressed tumor growth in athymic mice, whereas YAP1-LV cells formed tumors more aggressively than did control cells (3 mice with tumors in both flanks per each group). Error bars: mean ± SEM. *P<0.05, **P<0.001.

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1) Product Images from "Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy"

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy

Journal: Molecular cancer therapeutics

doi: 10.1158/1535-7163.MCT-20-0024

Attenuation of YAP1 expression impairs cell proliferation and increases sensitivity to anti-cancer agents in LUAD. A, Western blotting confirmed YAP1-knockdown in H1299 and H1437 cells and YAP1-overexpression in A549 cells. B, Proliferation assay: YAP1-deficient cells (H1299 YAP1-sh and H1437 YAP1-sh) showed lower proliferative ratios than did control cells, whereas YAP1-overexpressing cells (A549 YAP1-LV) proliferated more rapidly. C, Cells were treated with CDDP, and cell viability was recorded at 72 hours after treatment. H1299 and H1437 YAP1-sh cells showed greater sensitivity to CDDP, and YAP1-LV cells showed higher resistance, than did their control counterparts. D, YAP1-deficient cells treated with CDDP (2 μM or 5 μM for 72 hours) showed greater percentages of cells undergoing apoptosis than did control cells; whereas YAP1-LV cells showed substantially less apoptosis. E, Both YAP1-sh cell lines showed significantly suppressed tumor growth in athymic mice, whereas YAP1-LV cells formed tumors more aggressively than did control cells (3 mice with tumors in both flanks per each group). Error bars: mean ± SEM. *P<0.05, **P<0.001.

Figure Legend Snippet: Attenuation of YAP1 expression impairs cell proliferation and increases sensitivity to anti-cancer agents in LUAD. A, Western blotting confirmed YAP1-knockdown in H1299 and H1437 cells and YAP1-overexpression in A549 cells. B, Proliferation assay: YAP1-deficient cells (H1299 YAP1-sh and H1437 YAP1-sh) showed lower proliferative ratios than did control cells, whereas YAP1-overexpressing cells (A549 YAP1-LV) proliferated more rapidly. C, Cells were treated with CDDP, and cell viability was recorded at 72 hours after treatment. H1299 and H1437 YAP1-sh cells showed greater sensitivity to CDDP, and YAP1-LV cells showed higher resistance, than did their control counterparts. D, YAP1-deficient cells treated with CDDP (2 μM or 5 μM for 72 hours) showed greater percentages of cells undergoing apoptosis than did control cells; whereas YAP1-LV cells showed substantially less apoptosis. E, Both YAP1-sh cell lines showed significantly suppressed tumor growth in athymic mice, whereas YAP1-LV cells formed tumors more aggressively than did control cells (3 mice with tumors in both flanks per each group). Error bars: mean ± SEM. *P<0.05, **P<0.001.

Techniques Used: Expressing, Western Blot, Knockdown, Over Expression, Proliferation Assay, Control

YAP1 promotes STAT3 phosphorylation via IL-6 upregulation. A, Immunoblotting shows YAP1, STAT3 and pSTAT3 expressions in YAP1-deficient (H1299 and H1437) and YAP1-overexpressing (A549) LUAD cells. YAP1 expression was positively associated with pSTAT3 expression. B, qRT-PCR data show IL-6 mRNA expression to be positively associated with YAP1 expression in H1299, H1437 and A549 cell lines. C, Secretion of IL-6 into media was significantly reduced in YAP1-sh cells, whereas higher levels were observed in YAP1-LV cells, as measured by ELISA in cell culture supernatant. Immunoblotting of pSTAT3 and β-actin in the respective cell lines are shown below the graph. D, Addition of recombinant human IL-6 (0.1, 1 or 10 ng/ml) to H1299 and H1437 YAP1-sh cells increased pSTAT3 levels dose-dependently. E, Blocking IL-6 activity with IL-6 neutralizing antibody (1 μg/ml) inhibited pSTAT3 in a time-dependent manner without influencing total STAT3 expression in A549 YAP1-LV cells. F, ChIP-PCR assay with YAP1 antibody was conducted using H1299, H1437 and A549 cells. DNA isolation followed by PCR showed that YAP1 protein directly binds to the IL-6 gene promoter region in three LUAD cells analyzed (white arrows). “Input” indicates DNA lysate before ChIP, which was diluted to 2% for PCR reactions. Histone H3 antibody and normal rabbit IgG served as positive and negative control, respectively. G, Schematic shows YAP1 binding to IL-6 promoter and upregulating its transcription to induce STAT3 phosphorylation. Error bars: mean ± SEM. *P<0.05, **P<0.001.

Figure Legend Snippet: YAP1 promotes STAT3 phosphorylation via IL-6 upregulation. A, Immunoblotting shows YAP1, STAT3 and pSTAT3 expressions in YAP1-deficient (H1299 and H1437) and YAP1-overexpressing (A549) LUAD cells. YAP1 expression was positively associated with pSTAT3 expression. B, qRT-PCR data show IL-6 mRNA expression to be positively associated with YAP1 expression in H1299, H1437 and A549 cell lines. C, Secretion of IL-6 into media was significantly reduced in YAP1-sh cells, whereas higher levels were observed in YAP1-LV cells, as measured by ELISA in cell culture supernatant. Immunoblotting of pSTAT3 and β-actin in the respective cell lines are shown below the graph. D, Addition of recombinant human IL-6 (0.1, 1 or 10 ng/ml) to H1299 and H1437 YAP1-sh cells increased pSTAT3 levels dose-dependently. E, Blocking IL-6 activity with IL-6 neutralizing antibody (1 μg/ml) inhibited pSTAT3 in a time-dependent manner without influencing total STAT3 expression in A549 YAP1-LV cells. F, ChIP-PCR assay with YAP1 antibody was conducted using H1299, H1437 and A549 cells. DNA isolation followed by PCR showed that YAP1 protein directly binds to the IL-6 gene promoter region in three LUAD cells analyzed (white arrows). “Input” indicates DNA lysate before ChIP, which was diluted to 2% for PCR reactions. Histone H3 antibody and normal rabbit IgG served as positive and negative control, respectively. G, Schematic shows YAP1 binding to IL-6 promoter and upregulating its transcription to induce STAT3 phosphorylation. Error bars: mean ± SEM. *P<0.05, **P<0.001.

Techniques Used: Phospho-proteomics, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant, Blocking Assay, Activity Assay, DNA Extraction, Negative Control, Binding Assay

Positive correlation between YAP1 and pSTAT3 expressions in human LUAD tissues. A, Representative images of YAP1 and pSTAT3 immunostaining in the TMA (scale bar: 100 μm). B, IHC results indicate a statistically significant correlation between YAP1 and pSTAT3 expressions (P<0.0001, χ2 test). C, YAP1 and pSTAT3 expressions were evaluated by immunoblotting in 13 LUAD PDXs. All YAP1-expressing PDXs (CTG0162, 0178, 0502, 0848, 1309, 1762, 2017 and 2708) also expressed pSTAT3.

Figure Legend Snippet: Positive correlation between YAP1 and pSTAT3 expressions in human LUAD tissues. A, Representative images of YAP1 and pSTAT3 immunostaining in the TMA (scale bar: 100 μm). B, IHC results indicate a statistically significant correlation between YAP1 and pSTAT3 expressions (P<0.0001, χ2 test). C, YAP1 and pSTAT3 expressions were evaluated by immunoblotting in 13 LUAD PDXs. All YAP1-expressing PDXs (CTG0162, 0178, 0502, 0848, 1309, 1762, 2017 and 2708) also expressed pSTAT3.

Techniques Used: Immunostaining, Western Blot, Expressing

Evaluation of verteporfin and S3I-201 as inhibitors of YAP1 and STAT3, respectively, in LUAD cells. A, Verteporfin suppressed YAP1 and STAT3 expressions in H1299 and H1437 cells in a concentration-dependent manner. STAT3 monomer (black arrows) decreased as verteporfin concentration was increased, whereas high molecular weight complexes (regions surrounded by circles) increased, indicating oligomerization of STAT3. B, S3I-201 suppressed pSTAT3 in a dose-dependent manner.

Figure Legend Snippet: Evaluation of verteporfin and S3I-201 as inhibitors of YAP1 and STAT3, respectively, in LUAD cells. A, Verteporfin suppressed YAP1 and STAT3 expressions in H1299 and H1437 cells in a concentration-dependent manner. STAT3 monomer (black arrows) decreased as verteporfin concentration was increased, whereas high molecular weight complexes (regions surrounded by circles) increased, indicating oligomerization of STAT3. B, S3I-201 suppressed pSTAT3 in a dose-dependent manner.

Techniques Used: Concentration Assay, High Molecular Weight

Therapeutic efficacy of verteporfin and S3I-201 in cell lines and patient-derived preclinical xenografts mouse models of human LUAD. NSG mice were subcutaneously inoculated LUAD cell lines (H1299 and H1437 cells) and PDXs that endogenously expressed YAP1 and pSTAT3. Animals were randomly divided into different treatment groups (n=5) (A-D). Treatments were started when tumors reached 200 ± 50 mm3. A, Combined verteporfin + S3I-201 significantly inhibited tumor growth in a H1299 xenograft model. Verteporfin or S3I-201 treatment alone led to inhibited growth, but combined treatment had a greater inhibitory effect. B, In H1437 xenografts, only the combination of verteporfin + S3I-201 inhibited growth significantly more than controls, whereas either agent individually did not significantly affect tumor growth. C, D, Two EGFR wild-type PDXs (CTG0162 and CTG0178) that expressed both YAP1 and pSTAT3 (Fig. 3C) were implanted. The “All” combination (CDDP + GEM + verteporfin + S3I-201) dramatically impaired growth of both PDXs. Chemo + verteporfin also significantly retarded tumor growth until Day 28 in CTG0162 (C) and Day 34 in CTG0178 (D). However, after 4–5 weeks, tumors treated with chemo + verteporfin showed tumor regrowth. E, F, Pharmacodynamic analysis of PDX tumors treated with verteporfin, S3I-201 and chemotherapy drugs. Left panel: CTG0162 (E) and CTG0178 (F) tumors resected at 18 days after treatment (one sample per treatment). Both YAP1 and STAT3 were suppressed in the chemo + verteporfin and “All” treatment groups. Adding verteporfin to chemotherapy decreased NANOG and SOX2 expressions. Right panel: tumors resected at 43 days in CTG0162 (E) and 46 days in CTG0178 (F) were assessed (two samples per treatment). Expressions of YAP1 and STAT3 were higher in tumors treated with chemo + verteporfin than with the “All” group. Tumors from the “All” group had lower expression of NANOG in CTG0162, and both NANOG and SOX2 in CTG0178, than did the chemo + verteporfin group. Error bars: mean ± SEM. *P<0.05, **P<0.001. N.S., not significant.

Figure Legend Snippet: Therapeutic efficacy of verteporfin and S3I-201 in cell lines and patient-derived preclinical xenografts mouse models of human LUAD. NSG mice were subcutaneously inoculated LUAD cell lines (H1299 and H1437 cells) and PDXs that endogenously expressed YAP1 and pSTAT3. Animals were randomly divided into different treatment groups (n=5) (A-D). Treatments were started when tumors reached 200 ± 50 mm3. A, Combined verteporfin + S3I-201 significantly inhibited tumor growth in a H1299 xenograft model. Verteporfin or S3I-201 treatment alone led to inhibited growth, but combined treatment had a greater inhibitory effect. B, In H1437 xenografts, only the combination of verteporfin + S3I-201 inhibited growth significantly more than controls, whereas either agent individually did not significantly affect tumor growth. C, D, Two EGFR wild-type PDXs (CTG0162 and CTG0178) that expressed both YAP1 and pSTAT3 (Fig. 3C) were implanted. The “All” combination (CDDP + GEM + verteporfin + S3I-201) dramatically impaired growth of both PDXs. Chemo + verteporfin also significantly retarded tumor growth until Day 28 in CTG0162 (C) and Day 34 in CTG0178 (D). However, after 4–5 weeks, tumors treated with chemo + verteporfin showed tumor regrowth. E, F, Pharmacodynamic analysis of PDX tumors treated with verteporfin, S3I-201 and chemotherapy drugs. Left panel: CTG0162 (E) and CTG0178 (F) tumors resected at 18 days after treatment (one sample per treatment). Both YAP1 and STAT3 were suppressed in the chemo + verteporfin and “All” treatment groups. Adding verteporfin to chemotherapy decreased NANOG and SOX2 expressions. Right panel: tumors resected at 43 days in CTG0162 (E) and 46 days in CTG0178 (F) were assessed (two samples per treatment). Expressions of YAP1 and STAT3 were higher in tumors treated with chemo + verteporfin than with the “All” group. Tumors from the “All” group had lower expression of NANOG in CTG0162, and both NANOG and SOX2 in CTG0178, than did the chemo + verteporfin group. Error bars: mean ± SEM. *P<0.05, **P<0.001. N.S., not significant.

Techniques Used: Drug discovery, Derivative Assay, Expressing

Expressing:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at − 80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO 2 at 37°C.Cells were stored at − 80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO 2 at 37°C.

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Western Blot:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Knockdown:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Over Expression:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Proliferation Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Control:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Phospho-proteomics:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Quantitative RT-PCR:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Enzyme-linked Immunosorbent Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Cell Culture:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Recombinant:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Blocking Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Activity Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

DNA Extraction:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Negative Control:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Binding Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Immunostaining:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Concentration Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

High Molecular Weight:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Drug discovery:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Derivative Assay:

Article Title: Concurrent Targeting of Potential Cancer Stem Cells Regulating Pathways Sensitizes Lung Adenocarcinoma to Standard Chemotherapy
Article Snippet: EGFR wild-type LUAD cell lines (A549, H1299 and H1437) were obtained from ATCC (Manassas, VA, USA).. Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.Cells were stored at −80°C and cultured in RPMI-1640 (Mediatech, Manassas, VA, USA) for H1299 and H1437 and F-12K (Mediatech) for A549, supplemented with 10% fetal bovine serum (Atlanta Biologicals, Flowery Branch, GA, USA) in an atmosphere containing 5% CO2 at 37°C.

Review