canertinib Search Results


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  • 93
    Millipore canertinib
    Effect of <t>pan-ErbB</t> inhibitors on NRG-1 action on human melanocytes and MelanoDerms. ( A ) HEM-DP were treated for 9 days with 50 ng/ml NRG-1 in the presence of <t>C39</t> (0.5 μM) or CI-1033 (2 μM) or vehicle (DMSO) alone. The results are the average
    Canertinib, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Pfizer Inc canertinib
    Small molecule ErbB2 inhibitors increase sensitivity to BCR/ABL-directed TKI. Z119 cells were treated with indicated doses <t>canertinib</t> or lapatinib and (A) imatinib, (B) nilotinib or (C) dasatinib for 72 hours. Cells were stained with PI and the subdiploid population was measured by flow cytometry. Bars indicate the mean and SEM of at least three independent experiments.
    Canertinib, supplied by Pfizer Inc, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    LC Laboratories canertinib
    Growth factors overcome RTK-inhibited-phosphorylation of downstream targets. Examples of immunoblots showing effects of growth factors (100 ng/ml) on Akt and Erk phosphorylation (p) after sorafenib ( A ), dasatinib ( B ), <t>canertinib</t> ( C ), crizotinib ( D ) in pediatric low grade astrocytoma and ependymoma cells (2h). Controls represent the specific cell lines treated with DMSO. Growth-factor-driven rescue on cell viability as given in Fig. 3C and D is indicated underneath the blots: purple squares, no rescue; yellow/orange squares, partial rescue; red squares, complete rescue. Protein expression was normalized to β-actin as loading control and derived from three independent experiments of which the mean (± SEM) was calculated. EGF and HGF-driven rescue resulted in significantly higher protein expression of Akt (p = 0.028) and Erk (p = 0.043) during sorafenib in both UW-467 and Res-196 cells and during dasatinib in the low grade astrocytoma cell lines (p = 0.028 and p = 0.043). Significantly higher Akt phosphorylation was found after HGF addition during canertinib in Res-186, Res-259 and Res-196 cells (p = 0.043) of which Res-186 is shown as an example. *Discrepancies between growth-factor-driven rescue on cell viability level and downstream phosphorylation status.
    Canertinib, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 123 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Selleck Chemicals canertinib
    HPV8E6 directly binds to the ubiquitinated EGFR in an UV-dependent manner. (A) RTS3b cells were transiently transfected with a vector directing the expression of FLAG-tagged HPV8E6 under control of the CMV-promoter or the empty vector. Where indicated the cells have been irradiated with UV-light 30 min prior harvesting. 400 μg of extracts were incubated with the anti-EGFR-antibody (lanes 1–4) or with the FLAG-antibody (lanes 5–8), both coupled to sepharose, washed four times in 0.3 M KCl-buffer prior analysis by WB and 40 μg of the extracts were used in the input blots. (B) IP with anti-EGFR sepharose and 800 μg of extracts derived from the stable RTS3b-pLXSN and RTS3b-HPV8E6 lines, either left untreated or UV-irradiated 30 min prior harvesting. FLAG-HPV8E6 and EGFR present in the precipitate as well as in the input were detected by WB. (C) Extracts from UV-irradiated RTS3b cells that have been transiently transfected with the FLAG-HPV8E6 or the FLAG-vector were incubated either with DMSO (lanes 1, 2, 5), 3 μM (lane 3) or 7 μM (lane 4) <t>Canertinib</t> for 30 min. The IP was performed with the FLAG-antibody. All WB were developed with the indicated antibodies. In lanes 6–10, C33A cells were transiently transfected with expression vectors for EGFR-GFP and FLAG-HPV8E6, and UV-irradiated, where indicated, prior harvesting 30 min later. EGFR-GFP was precipitated from 400 μg of extract by the EGFR antibody and bound HPV8E6 was detected by the FLAG-antibody. The input blots were developed with the antibodies against pEGFR-Y1068, GFP, and FLAG. The positions of the molecular weight markers are provided (in kDa).
    Canertinib, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    LC Laboratories canertinib dihydrochloride salt
    APAP caused rapid mitochondrial translocation of EGFR and mitochondrial dysfunction, which was inhibited by EGFRi. A, Western blot analysis of phospho-EGFR and EGFR in mitochondrial fraction at 1.5 h after administration of APAP (300 mg/kg) in mice pretreated (2-h prior to APAP) with <t>canertinib</t> (80 mg/kg). B-E , Oxygen consumption rate analysis in freshly isolated mitochondria from mice ( n = 3) treated with APAP (300 mg/kg) + PBS, APAP (300 mg/kg) + canertinib (80 mg/kg) or saline (control), using Seahorse extracellular flux analyzer. B and C, Canertinib was administered 2hr before APAP and analysis was done 1.5 h after APAP treatment. D and E, Canertinib was administered 1-h post-APAP and analysis was done 3 h after APAP treatment. Oxidative phosphorylation was manipulated with injection of ADP, oligomycin (oligo), FCCP, and antimycin A (anti-A). Respiration was sequentially measured in a coupled state with substrate present (basal respiration), followed by State 3 (phosphorylating respiration, in the presence of ADP and substrate), State 4o (leak state induced with the addition of oligomycin - inhibitor of ATP synthase), and then maximal uncoupler (FCCP)-stimulated respiration (State 3u). At the end, antimycin A (complex III inhibitor) was added to inhibit mitochondrial respiration completely * indicate significant difference w.r.t. to APAP + PBS group at P
    Canertinib Dihydrochloride Salt, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Warner-Lambert canertinib
    APAP caused rapid mitochondrial translocation of EGFR and mitochondrial dysfunction, which was inhibited by EGFRi. A, Western blot analysis of phospho-EGFR and EGFR in mitochondrial fraction at 1.5 h after administration of APAP (300 mg/kg) in mice pretreated (2-h prior to APAP) with <t>canertinib</t> (80 mg/kg). B-E , Oxygen consumption rate analysis in freshly isolated mitochondria from mice ( n = 3) treated with APAP (300 mg/kg) + PBS, APAP (300 mg/kg) + canertinib (80 mg/kg) or saline (control), using Seahorse extracellular flux analyzer. B and C, Canertinib was administered 2hr before APAP and analysis was done 1.5 h after APAP treatment. D and E, Canertinib was administered 1-h post-APAP and analysis was done 3 h after APAP treatment. Oxidative phosphorylation was manipulated with injection of ADP, oligomycin (oligo), FCCP, and antimycin A (anti-A). Respiration was sequentially measured in a coupled state with substrate present (basal respiration), followed by State 3 (phosphorylating respiration, in the presence of ADP and substrate), State 4o (leak state induced with the addition of oligomycin - inhibitor of ATP synthase), and then maximal uncoupler (FCCP)-stimulated respiration (State 3u). At the end, antimycin A (complex III inhibitor) was added to inhibit mitochondrial respiration completely * indicate significant difference w.r.t. to APAP + PBS group at P
    Canertinib, supplied by Warner-Lambert, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    BioVision pan erbb inhibitor ci 1033
    Modulation of PI3-K activity inversely impacts InlB-requirement for Lm entry in vitro. (A) Whole cell extracts from LS174T cells incubated with PBS or different concentrations of the <t>ErbB</t> inhibitor <t>CI-1033</t> (0.01, 0.1, or 1 µM) during 24 h, immunoblotted for P-Akt and total Akt. (B) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in LS174T cells incubated with PBS (controls, black bars) or CI-1033 1 µM (gray bars) for 24 h. (C) Whole cell extracts from LS174T cells incubated with PBS or CI-1033 1 µM and infected with WT Lm EGD and isogenic mutant Δ inlB , immunoblotted for P-Akt and total Akt. (D) Whole cell extracts from Jar cells incubated with PBS or EGF 50 ng/ml during 10, 30, or 60 min, immunoblotted for P-Akt and total Akt. (E) Whole cell extracts from Jar cells transfected with esiRNA PTEN, immunoblotted for PTEN. (F) Whole cell extracts from Jar cells transfected with esiRNA PTEN or control scrambled and incubated with PBS or EGF 50 ng/ml for 120 min, immunoblotted for P-Akt and total Akt. (G) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS, or transfected with esiRNA PTEN or incubated with EGF 50 ng/ml. (H) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS or incubated with EGF 50 ng/ml, and transfected with esiRNA PTEN. (B, G, and H) Invasion assays were performed in triplicates and represent at least three independent experiments for each condition tested. Relative entry represents the ratio of CFUs for each strain divided by the mean of CFUs for the WT Lm EGD strain. A one-way ANOVA test followed by a Bonferroni’s multiple comparisons was performed. Error bars, SEM. (A, C, D, E, and F) Immunoblots were performed at least two times. Densitometry was performed and the ratio of P-Akt over total Akt is relative to the PBS, for which the value is normalized to 1.
    Pan Erbb Inhibitor Ci 1033, supplied by BioVision, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    LC Laboratories pan erbb inhibitor canertinib
    Modulation of PI3-K activity inversely impacts InlB-requirement for Lm entry in vitro. (A) Whole cell extracts from LS174T cells incubated with PBS or different concentrations of the <t>ErbB</t> inhibitor <t>CI-1033</t> (0.01, 0.1, or 1 µM) during 24 h, immunoblotted for P-Akt and total Akt. (B) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in LS174T cells incubated with PBS (controls, black bars) or CI-1033 1 µM (gray bars) for 24 h. (C) Whole cell extracts from LS174T cells incubated with PBS or CI-1033 1 µM and infected with WT Lm EGD and isogenic mutant Δ inlB , immunoblotted for P-Akt and total Akt. (D) Whole cell extracts from Jar cells incubated with PBS or EGF 50 ng/ml during 10, 30, or 60 min, immunoblotted for P-Akt and total Akt. (E) Whole cell extracts from Jar cells transfected with esiRNA PTEN, immunoblotted for PTEN. (F) Whole cell extracts from Jar cells transfected with esiRNA PTEN or control scrambled and incubated with PBS or EGF 50 ng/ml for 120 min, immunoblotted for P-Akt and total Akt. (G) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS, or transfected with esiRNA PTEN or incubated with EGF 50 ng/ml. (H) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS or incubated with EGF 50 ng/ml, and transfected with esiRNA PTEN. (B, G, and H) Invasion assays were performed in triplicates and represent at least three independent experiments for each condition tested. Relative entry represents the ratio of CFUs for each strain divided by the mean of CFUs for the WT Lm EGD strain. A one-way ANOVA test followed by a Bonferroni’s multiple comparisons was performed. Error bars, SEM. (A, C, D, E, and F) Immunoblots were performed at least two times. Densitometry was performed and the ratio of P-Akt over total Akt is relative to the PBS, for which the value is normalized to 1.
    Pan Erbb Inhibitor Canertinib, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    LC Laboratories canertinib dihydrochloride
    Modulation of PI3-K activity inversely impacts InlB-requirement for Lm entry in vitro. (A) Whole cell extracts from LS174T cells incubated with PBS or different concentrations of the <t>ErbB</t> inhibitor <t>CI-1033</t> (0.01, 0.1, or 1 µM) during 24 h, immunoblotted for P-Akt and total Akt. (B) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in LS174T cells incubated with PBS (controls, black bars) or CI-1033 1 µM (gray bars) for 24 h. (C) Whole cell extracts from LS174T cells incubated with PBS or CI-1033 1 µM and infected with WT Lm EGD and isogenic mutant Δ inlB , immunoblotted for P-Akt and total Akt. (D) Whole cell extracts from Jar cells incubated with PBS or EGF 50 ng/ml during 10, 30, or 60 min, immunoblotted for P-Akt and total Akt. (E) Whole cell extracts from Jar cells transfected with esiRNA PTEN, immunoblotted for PTEN. (F) Whole cell extracts from Jar cells transfected with esiRNA PTEN or control scrambled and incubated with PBS or EGF 50 ng/ml for 120 min, immunoblotted for P-Akt and total Akt. (G) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS, or transfected with esiRNA PTEN or incubated with EGF 50 ng/ml. (H) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS or incubated with EGF 50 ng/ml, and transfected with esiRNA PTEN. (B, G, and H) Invasion assays were performed in triplicates and represent at least three independent experiments for each condition tested. Relative entry represents the ratio of CFUs for each strain divided by the mean of CFUs for the WT Lm EGD strain. A one-way ANOVA test followed by a Bonferroni’s multiple comparisons was performed. Error bars, SEM. (A, C, D, E, and F) Immunoblots were performed at least two times. Densitometry was performed and the ratio of P-Akt over total Akt is relative to the PBS, for which the value is normalized to 1.
    Canertinib Dihydrochloride, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    AK Scientific canertinib
    Validation of selected GenEx-TNBC predictions in TNBC cell lines of multiple molecular subtypes Crystal violet staining of HCC1937 (BL1), BT549 (ML), and MDA-MB-453 (LAR) cells grown in the presence of the indicated concentrations of Mecamylamine (A) , <t>Canertinib</t> (B) , Bortezomib (C) , and Tretinoin (D) for seven (7) days. Data for each cell line were normalized to the appropriate solvent control (ethanol for Mecamylamine, DMSO for all others). Data were analyzed by two-way ANOVA with Tukey post hoc multiple comparisons test (asterisks indicate per-dose comparisons) and are presented as the mean +/− standard deviation (S.D.) for a single experiment performed with 6 technical replicates that is representative of at least two independent experiments (biological replicates).
    Canertinib, supplied by AK Scientific, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Axon Medchem LLC canertinib
    Validation of selected GenEx-TNBC predictions in TNBC cell lines of multiple molecular subtypes Crystal violet staining of HCC1937 (BL1), BT549 (ML), and MDA-MB-453 (LAR) cells grown in the presence of the indicated concentrations of Mecamylamine (A) , <t>Canertinib</t> (B) , Bortezomib (C) , and Tretinoin (D) for seven (7) days. Data for each cell line were normalized to the appropriate solvent control (ethanol for Mecamylamine, DMSO for all others). Data were analyzed by two-way ANOVA with Tukey post hoc multiple comparisons test (asterisks indicate per-dose comparisons) and are presented as the mean +/− standard deviation (S.D.) for a single experiment performed with 6 technical replicates that is representative of at least two independent experiments (biological replicates).
    Canertinib, supplied by Axon Medchem LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    LC Laboratories egf canertinib
    Validation of selected GenEx-TNBC predictions in TNBC cell lines of multiple molecular subtypes Crystal violet staining of HCC1937 (BL1), BT549 (ML), and MDA-MB-453 (LAR) cells grown in the presence of the indicated concentrations of Mecamylamine (A) , <t>Canertinib</t> (B) , Bortezomib (C) , and Tretinoin (D) for seven (7) days. Data for each cell line were normalized to the appropriate solvent control (ethanol for Mecamylamine, DMSO for all others). Data were analyzed by two-way ANOVA with Tukey post hoc multiple comparisons test (asterisks indicate per-dose comparisons) and are presented as the mean +/− standard deviation (S.D.) for a single experiment performed with 6 technical replicates that is representative of at least two independent experiments (biological replicates).
    Egf Canertinib, supplied by LC Laboratories, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    Potent EGFR kinase inhibitor
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    N/A
    The HER family of receptor tyrosine kinases EGFR HER2 HER3 and HER4 mediate proliferation migration adhesion differentiation and survival in many different cell types and have been implicated in the
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    Image Search Results


    Effect of pan-ErbB inhibitors on NRG-1 action on human melanocytes and MelanoDerms. ( A ) HEM-DP were treated for 9 days with 50 ng/ml NRG-1 in the presence of C39 (0.5 μM) or CI-1033 (2 μM) or vehicle (DMSO) alone. The results are the average

    Journal: Journal of Cell Science

    Article Title: The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin

    doi: 10.1242/jcs.064774

    Figure Lengend Snippet: Effect of pan-ErbB inhibitors on NRG-1 action on human melanocytes and MelanoDerms. ( A ) HEM-DP were treated for 9 days with 50 ng/ml NRG-1 in the presence of C39 (0.5 μM) or CI-1033 (2 μM) or vehicle (DMSO) alone. The results are the average

    Article Snippet: For the experiments where the MelanoDerms were treated with NRG-1 and/or various pan-ErbB inhibitors, the dark MelanoDerms were first incubated with a pan-ErbB inhibitor (0.5 μM C39, EMD Chemicals, Gibbistown, NJ or 2 μM CI-1033, Selleck Chemicals LLC, US) for 30 minutes or with vehicle (DMSO) only before treatment with 50 ng/ml NRG-1; identical amounts of DMSO (0.02%) were added to all samples.

    Techniques:

    Measuring of EGFR and c-erbB2 expression; activation of ERK1/2 and AKT in MCF-7, T47D, and their tamoxifen-resistant derivative cells MCF-7(TamR) and T47D (TamR) cells, respectively. (a) Changes in levels of total EGFR and c-erbB2 expression by Western blot. (b) Changes in levels of p-EGFR and p-c-erbB2 expression by Western blot. (c) Changes in levels of expression and activation of ERK1/2 and AKT by Western blot. (d) Changes in levels of EGFR expression and activation in t47d cells before and after the development of tamoxifen resistance by Western blot. (e) The inhibitory effects of canertinib on the activation of EGFR, Akt, and ERK1/2 and their encoded protein in both MCF-7 and MCF-7(TamR) cells by Western blot.

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Measuring of EGFR and c-erbB2 expression; activation of ERK1/2 and AKT in MCF-7, T47D, and their tamoxifen-resistant derivative cells MCF-7(TamR) and T47D (TamR) cells, respectively. (a) Changes in levels of total EGFR and c-erbB2 expression by Western blot. (b) Changes in levels of p-EGFR and p-c-erbB2 expression by Western blot. (c) Changes in levels of expression and activation of ERK1/2 and AKT by Western blot. (d) Changes in levels of EGFR expression and activation in t47d cells before and after the development of tamoxifen resistance by Western blot. (e) The inhibitory effects of canertinib on the activation of EGFR, Akt, and ERK1/2 and their encoded protein in both MCF-7 and MCF-7(TamR) cells by Western blot.

    Article Snippet: Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Expressing, Activation Assay, Western Blot

    Chemical structure of canertinib (EGFR-irreversible tyrosine kinase inhibitor).

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Chemical structure of canertinib (EGFR-irreversible tyrosine kinase inhibitor).

    Article Snippet: Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques:

    Effect of EGFR inhibition with canertinib treatment on the sensitivity of MCF-7 and MCF-7(TamR) cells to a 48-hour treatment with (a) paclitaxel and (b) daunorubicin. Cells were subjected to a 48-hour cytotoxic treatment with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib before 4 days recovery in the same medium minus the cytotoxic. P values calculated from a paired t -test comparing growth inhibition between cells untreated with canertinib, both of which were treated with a given concentration of daunorubicin or paclitaxel. ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect of EGFR inhibition with canertinib treatment on the sensitivity of MCF-7 and MCF-7(TamR) cells to a 48-hour treatment with (a) paclitaxel and (b) daunorubicin. Cells were subjected to a 48-hour cytotoxic treatment with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib before 4 days recovery in the same medium minus the cytotoxic. P values calculated from a paired t -test comparing growth inhibition between cells untreated with canertinib, both of which were treated with a given concentration of daunorubicin or paclitaxel. ∗ indicates p

    Article Snippet: Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Inhibition, Concentration Assay

    Effect of EGFR inhibition with canertinib treatment on the sensitivity of (a) MCF-7 and (b) MCF-7(TamR) cells to carboplatin. Cells were subjected to a 9-day treatment with carboplatin with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib to the culture medium for the full length of the treatment, refreshed on day 5. P values calculated from a paired t -test comparing growth inhibition between cells treated with canertinib in addition to a given concentration of paclitaxel. ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect of EGFR inhibition with canertinib treatment on the sensitivity of (a) MCF-7 and (b) MCF-7(TamR) cells to carboplatin. Cells were subjected to a 9-day treatment with carboplatin with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib to the culture medium for the full length of the treatment, refreshed on day 5. P values calculated from a paired t -test comparing growth inhibition between cells treated with canertinib in addition to a given concentration of paclitaxel. ∗ indicates p

    Article Snippet: Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Inhibition, Concentration Assay

    Effect on growth of MCF-7 (red) and MCF-7(TamR) (blue) cells of a 9-day treatment of the EGFR inhibitors (a) canertinib and (b) canertinib added to the cell culture medium and refreshed on day 5 (test comparing growth inhibition between cell lines at a given concentration of EGFR inhibitor). ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect on growth of MCF-7 (red) and MCF-7(TamR) (blue) cells of a 9-day treatment of the EGFR inhibitors (a) canertinib and (b) canertinib added to the cell culture medium and refreshed on day 5 (test comparing growth inhibition between cell lines at a given concentration of EGFR inhibitor). ∗ indicates p

    Article Snippet: Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Cell Culture, Inhibition, Concentration Assay

    Profile of EGFR tyrosine phosphorylation in cells pre-incubated with antioxidants or EGFR tyrosine kinase inhibitor and exposed to NBD compounds. MDA MB468 cells were pre-incubated with 0,2% DMSO ( a ) 500 U/ml PEG-catalase ( b ), 5 mM NAC ( c ), 5 mM thioglycerol ( d ) or 2 μM CI-1033 ( e ) and exposed to NBD compounds (100 μM), EGF (100 ng/ml) or vehicle for 15 min.

    Journal: Scientific Reports

    Article Title: Activation of EGFR by small compounds through coupling the generation of hydrogen peroxide to stable dimerization of Cu/Zn SOD1

    doi: 10.1038/srep21088

    Figure Lengend Snippet: Profile of EGFR tyrosine phosphorylation in cells pre-incubated with antioxidants or EGFR tyrosine kinase inhibitor and exposed to NBD compounds. MDA MB468 cells were pre-incubated with 0,2% DMSO ( a ) 500 U/ml PEG-catalase ( b ), 5 mM NAC ( c ), 5 mM thioglycerol ( d ) or 2 μM CI-1033 ( e ) and exposed to NBD compounds (100 μM), EGF (100 ng/ml) or vehicle for 15 min.

    Article Snippet: Catalase, PEG-catalase, thioglycerol, NAC, and compound CI-1033 were from Sigma-Aldrich.

    Techniques: Incubation, Multiple Displacement Amplification

    Dimerization of SOD1 in cancer cells exposed to lipophilic NBD compounds, and SOD1 exposed to NBD compounds in vitro . ( a ) Western blot analysis of proteins in MDA MB468 cells pre-incubated with NAC (5 mM), PEG-catalase (500 U/ml), thioglycerol (5 mM) or CI-1033 (2 μM) for 30 min, and then exposed to NBD compounds (100 μM) or EGF (500 ng/ml) for 15 min. SOD1 monomers were present in all cells (shown only in cells pre-incubated with DMSO). ( b ) Detection of NBD compounds bound to purified human SOD1 with Coomassie staining. ( c) Fluorescence detection of NBD compound/SOD1 complexes blotted onto NC membrane using a Typhoon 9410 scanner; the excitation wavelength was 488 nm and the emission wavelength was 526 nm.

    Journal: Scientific Reports

    Article Title: Activation of EGFR by small compounds through coupling the generation of hydrogen peroxide to stable dimerization of Cu/Zn SOD1

    doi: 10.1038/srep21088

    Figure Lengend Snippet: Dimerization of SOD1 in cancer cells exposed to lipophilic NBD compounds, and SOD1 exposed to NBD compounds in vitro . ( a ) Western blot analysis of proteins in MDA MB468 cells pre-incubated with NAC (5 mM), PEG-catalase (500 U/ml), thioglycerol (5 mM) or CI-1033 (2 μM) for 30 min, and then exposed to NBD compounds (100 μM) or EGF (500 ng/ml) for 15 min. SOD1 monomers were present in all cells (shown only in cells pre-incubated with DMSO). ( b ) Detection of NBD compounds bound to purified human SOD1 with Coomassie staining. ( c) Fluorescence detection of NBD compound/SOD1 complexes blotted onto NC membrane using a Typhoon 9410 scanner; the excitation wavelength was 488 nm and the emission wavelength was 526 nm.

    Article Snippet: Catalase, PEG-catalase, thioglycerol, NAC, and compound CI-1033 were from Sigma-Aldrich.

    Techniques: In Vitro, Western Blot, Multiple Displacement Amplification, Incubation, Purification, Staining, Fluorescence

    Measuring of EGFR and c-erbB2 expression; activation of ERK1/2 and AKT in MCF-7, T47D, and their tamoxifen-resistant derivative cells MCF-7(TamR) and T47D (TamR) cells, respectively. (a) Changes in levels of total EGFR and c-erbB2 expression by Western blot. (b) Changes in levels of p-EGFR and p-c-erbB2 expression by Western blot. (c) Changes in levels of expression and activation of ERK1/2 and AKT by Western blot. (d) Changes in levels of EGFR expression and activation in t47d cells before and after the development of tamoxifen resistance by Western blot. (e) The inhibitory effects of canertinib on the activation of EGFR, Akt, and ERK1/2 and their encoded protein in both MCF-7 and MCF-7(TamR) cells by Western blot.

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Measuring of EGFR and c-erbB2 expression; activation of ERK1/2 and AKT in MCF-7, T47D, and their tamoxifen-resistant derivative cells MCF-7(TamR) and T47D (TamR) cells, respectively. (a) Changes in levels of total EGFR and c-erbB2 expression by Western blot. (b) Changes in levels of p-EGFR and p-c-erbB2 expression by Western blot. (c) Changes in levels of expression and activation of ERK1/2 and AKT by Western blot. (d) Changes in levels of EGFR expression and activation in t47d cells before and after the development of tamoxifen resistance by Western blot. (e) The inhibitory effects of canertinib on the activation of EGFR, Akt, and ERK1/2 and their encoded protein in both MCF-7 and MCF-7(TamR) cells by Western blot.

    Article Snippet: Materials Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Expressing, Activation Assay, Western Blot

    Chemical structure of canertinib (EGFR-irreversible tyrosine kinase inhibitor).

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Chemical structure of canertinib (EGFR-irreversible tyrosine kinase inhibitor).

    Article Snippet: Materials Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques:

    Effect of EGFR inhibition with canertinib treatment on the sensitivity of MCF-7 and MCF-7(TamR) cells to a 48-hour treatment with (a) paclitaxel and (b) daunorubicin. Cells were subjected to a 48-hour cytotoxic treatment with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib before 4 days recovery in the same medium minus the cytotoxic. P values calculated from a paired t -test comparing growth inhibition between cells untreated with canertinib, both of which were treated with a given concentration of daunorubicin or paclitaxel. ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect of EGFR inhibition with canertinib treatment on the sensitivity of MCF-7 and MCF-7(TamR) cells to a 48-hour treatment with (a) paclitaxel and (b) daunorubicin. Cells were subjected to a 48-hour cytotoxic treatment with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib before 4 days recovery in the same medium minus the cytotoxic. P values calculated from a paired t -test comparing growth inhibition between cells untreated with canertinib, both of which were treated with a given concentration of daunorubicin or paclitaxel. ∗ indicates p

    Article Snippet: Materials Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Inhibition, Concentration Assay

    Effect of EGFR inhibition with canertinib treatment on the sensitivity of (a) MCF-7 and (b) MCF-7(TamR) cells to carboplatin. Cells were subjected to a 9-day treatment with carboplatin with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib to the culture medium for the full length of the treatment, refreshed on day 5. P values calculated from a paired t -test comparing growth inhibition between cells treated with canertinib in addition to a given concentration of paclitaxel. ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect of EGFR inhibition with canertinib treatment on the sensitivity of (a) MCF-7 and (b) MCF-7(TamR) cells to carboplatin. Cells were subjected to a 9-day treatment with carboplatin with the addition of (red) 0 μ M, (blue) 0.1 μ M, or (green) 1 μ M canertinib to the culture medium for the full length of the treatment, refreshed on day 5. P values calculated from a paired t -test comparing growth inhibition between cells treated with canertinib in addition to a given concentration of paclitaxel. ∗ indicates p

    Article Snippet: Materials Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Inhibition, Concentration Assay

    Effect on growth of MCF-7 (red) and MCF-7(TamR) (blue) cells of a 9-day treatment of the EGFR inhibitors (a) canertinib and (b) canertinib added to the cell culture medium and refreshed on day 5 (test comparing growth inhibition between cell lines at a given concentration of EGFR inhibitor). ∗ indicates p

    Journal: International Journal of Genomics

    Article Title: The Effect of Canertinib on Sensitivity of Cytotoxic Drugs in Tamoxifen-Resistant Breast Cancer Cells In Vitro

    doi: 10.1155/2018/7628734

    Figure Lengend Snippet: Effect on growth of MCF-7 (red) and MCF-7(TamR) (blue) cells of a 9-day treatment of the EGFR inhibitors (a) canertinib and (b) canertinib added to the cell culture medium and refreshed on day 5 (test comparing growth inhibition between cell lines at a given concentration of EGFR inhibitor). ∗ indicates p

    Article Snippet: Materials Clinical grade canertinib (CI-1033) and cytotoxic drugs (paclitaxel, etoposide, vinorelbine, and daunorubicin) were purchased from Sigma Chemical Co. (Sigma-Aldrich, Egypt).

    Techniques: Cell Culture, Inhibition, Concentration Assay

    Small molecule ErbB2 inhibitors increase sensitivity to BCR/ABL-directed TKI. Z119 cells were treated with indicated doses canertinib or lapatinib and (A) imatinib, (B) nilotinib or (C) dasatinib for 72 hours. Cells were stained with PI and the subdiploid population was measured by flow cytometry. Bars indicate the mean and SEM of at least three independent experiments.

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: Small molecule ErbB2 inhibitors increase sensitivity to BCR/ABL-directed TKI. Z119 cells were treated with indicated doses canertinib or lapatinib and (A) imatinib, (B) nilotinib or (C) dasatinib for 72 hours. Cells were stained with PI and the subdiploid population was measured by flow cytometry. Bars indicate the mean and SEM of at least three independent experiments.

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: Staining, Flow Cytometry, Cytometry

    Decreased proliferation of ErbB2 + Ph + ALL cells is induced by ErbB inhibition. Z119 and Z181 cells were treated with indicated doses of (A) canertinib or (B) lapatinib and allowed to proliferate for 96 hours. Cell viability and total cell number were determined every 24 hours by trypan blue exclusion. Points indicate the mean and SEM of viable cell numbers in at least three independent experiments.

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: Decreased proliferation of ErbB2 + Ph + ALL cells is induced by ErbB inhibition. Z119 and Z181 cells were treated with indicated doses of (A) canertinib or (B) lapatinib and allowed to proliferate for 96 hours. Cell viability and total cell number were determined every 24 hours by trypan blue exclusion. Points indicate the mean and SEM of viable cell numbers in at least three independent experiments.

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: Inhibition

    Reverse phase protein analyses of ErbB2 + Ph + ALL with canertinib treatment. (A) Unsupervised clustering analyses were performed on RPPA data from Z181 or Z119 cells treated with 0.1–5 µM of canertinib for 18 hours. To generate heat maps, total protein and phosphoprotein levels were quantified by RPPA, and data were centered on the mean. Intensifying red color indicates increasing protein or phosphoprotein expression relative to the mean, black indicates the mean value, and intensifying green color indicates decreasing levels. (B) Supervised clustering showing relative changes in expression of pro-apoptotic proteins (Bim, cleaved PARP), protein kinase Cs, heat shock proteins, p38 mitogen activated protein kinase, and p38 T180/Y182p; five components of the phosphatidylinositol 3 kinase signaling pathway (Akt T308p, Akt S473p, p70S6-kinase T389p, S6-kinase S235/36p, and S6-kinase S240/44p); and ErbB2 Y1248p in the two cell lines after treatment.

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: Reverse phase protein analyses of ErbB2 + Ph + ALL with canertinib treatment. (A) Unsupervised clustering analyses were performed on RPPA data from Z181 or Z119 cells treated with 0.1–5 µM of canertinib for 18 hours. To generate heat maps, total protein and phosphoprotein levels were quantified by RPPA, and data were centered on the mean. Intensifying red color indicates increasing protein or phosphoprotein expression relative to the mean, black indicates the mean value, and intensifying green color indicates decreasing levels. (B) Supervised clustering showing relative changes in expression of pro-apoptotic proteins (Bim, cleaved PARP), protein kinase Cs, heat shock proteins, p38 mitogen activated protein kinase, and p38 T180/Y182p; five components of the phosphatidylinositol 3 kinase signaling pathway (Akt T308p, Akt S473p, p70S6-kinase T389p, S6-kinase S235/36p, and S6-kinase S240/44p); and ErbB2 Y1248p in the two cell lines after treatment.

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: Expressing

    Apoptosis of ErbB2 + Ph + ALL cell lines is induced by canertinib. (A and B, upper panels) Z119 and Z181 cells were treated with canertinib for 18 hours and lysates were subjected to SDS-PAGE followed by immunoblotting with the indicated antibodies. (A and B, lower panels) Protein expression values from RPPA analyses were quantified, and expression relative to the mean graphed. Triangles indicate drug doses of 0–5 µM. *p

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: Apoptosis of ErbB2 + Ph + ALL cell lines is induced by canertinib. (A and B, upper panels) Z119 and Z181 cells were treated with canertinib for 18 hours and lysates were subjected to SDS-PAGE followed by immunoblotting with the indicated antibodies. (A and B, lower panels) Protein expression values from RPPA analyses were quantified, and expression relative to the mean graphed. Triangles indicate drug doses of 0–5 µM. *p

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: SDS Page, Expressing

    ErbB2 protein expression and activation in Ph + ALL cell lines. (A) Two established human Ph + ALL cell lines, Z181 and Z119, were lysed and then subjected to SDS-PAGE followed by Western blotting for ErbB2 and actin. Blots are representative of at least three independent experiments. Densitometry was performed using ImageJ (National Institutes of Health). (B) Cell lines were stained with murine PE-conjugated anti-human ErbB2 monoclonal antibody and assessed by flow cytometry; isotype control staining (grey) and anti-ErbB2 staining (white). Cell surface quantification was performed as described in Materials and Methods. Numbers indicate the average number of molecules of ErbB2 per cell. (C) Protein lysates were collected from cells treated with the indicated doses of canertinib for 18 hours. Samples were subjected to SDS-PAGE followed by Western blotting for ErbB2 Y1248p (ErbB2p), total ErbB2 (ErbB2), or actin. Blots are representative of at least three experiments. Densitometry was performed using ImageJ software and normalized to actin. (D) RPPA analyses were performed utilizing ErbB2p antibody. Bars represent the means of three individual experiments. Triangles indicate drug concentrations of 0–5 µM. *p

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: ErbB2 protein expression and activation in Ph + ALL cell lines. (A) Two established human Ph + ALL cell lines, Z181 and Z119, were lysed and then subjected to SDS-PAGE followed by Western blotting for ErbB2 and actin. Blots are representative of at least three independent experiments. Densitometry was performed using ImageJ (National Institutes of Health). (B) Cell lines were stained with murine PE-conjugated anti-human ErbB2 monoclonal antibody and assessed by flow cytometry; isotype control staining (grey) and anti-ErbB2 staining (white). Cell surface quantification was performed as described in Materials and Methods. Numbers indicate the average number of molecules of ErbB2 per cell. (C) Protein lysates were collected from cells treated with the indicated doses of canertinib for 18 hours. Samples were subjected to SDS-PAGE followed by Western blotting for ErbB2 Y1248p (ErbB2p), total ErbB2 (ErbB2), or actin. Blots are representative of at least three experiments. Densitometry was performed using ImageJ software and normalized to actin. (D) RPPA analyses were performed utilizing ErbB2p antibody. Bars represent the means of three individual experiments. Triangles indicate drug concentrations of 0–5 µM. *p

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: Expressing, Activation Assay, SDS Page, Western Blot, Staining, Flow Cytometry, Cytometry, Software

    Downregulation of pro-survival signaling induced by canertinib. (A) Protein expression values from RPPA analyses of Z119 (white) and Z181 (black) cells were quantified, and expression relative to the mean graphed. Triangles indicate drug concentrations of 0–5 µM. *p

    Journal: PLoS ONE

    Article Title: Small Molecule ErbB Inhibitors Decrease Proliferative Signaling and Promote Apoptosis in Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia

    doi: 10.1371/journal.pone.0070608

    Figure Lengend Snippet: Downregulation of pro-survival signaling induced by canertinib. (A) Protein expression values from RPPA analyses of Z119 (white) and Z181 (black) cells were quantified, and expression relative to the mean graphed. Triangles indicate drug concentrations of 0–5 µM. *p

    Article Snippet: Canertinib was received from Pfizer, Inc. (New York, NY) and lapatinib, imatinib, nilotinib, and dasatinib were purchased from LC Laboratories (Woburn, MA).

    Techniques: Expressing

    Drug elution of T. brucei protein kinases using an ATP-affinity resin. Proteins in total cell lysate from T. brucei were bound on ATP resin. Sepharose 4B resin was used as control. Unbound proteins (Flow through) were recovered and resin was washed sequentially with buffer A and buffer A containing 1 M KCl (Panel A). Bound proteins were eluted with lapatinib (100 µM), or AEE788 (100 µM) or canertinib (100 µM) (Panel B, C and D, respectively). Proteins were visualized by silver staining. Matrix label, A is ATP-sepharose, and S is Sepharose 4B.

    Journal: PLoS ONE

    Article Title: Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds

    doi: 10.1371/journal.pone.0056150

    Figure Lengend Snippet: Drug elution of T. brucei protein kinases using an ATP-affinity resin. Proteins in total cell lysate from T. brucei were bound on ATP resin. Sepharose 4B resin was used as control. Unbound proteins (Flow through) were recovered and resin was washed sequentially with buffer A and buffer A containing 1 M KCl (Panel A). Bound proteins were eluted with lapatinib (100 µM), or AEE788 (100 µM) or canertinib (100 µM) (Panel B, C and D, respectively). Proteins were visualized by silver staining. Matrix label, A is ATP-sepharose, and S is Sepharose 4B.

    Article Snippet: AEE788 (Novartis) inhibits EGFR and VEGFR , , and canertinib (CI-1033) (Pfizer) is a pan-inhibitor of EGFRs – .

    Techniques: Flow Cytometry, Silver Staining

    Lapatinib, AEE788 and canertinib kill bloodstream T. brucei . Bloodstream form T. brucei (initial cell density of 2×10 3 cells/ml) were cultured in 24-well plates for 48 h with either DMSO (control) or different concentrations of drug. Cells were counted with a hemocytometer and the graphs plotted. Data are mean ± standard deviation obtained from two independent experiments performed in duplicate. ( A ). Effect of lapatinib on growth of T. brucei . ( B ). Comparison of growth inhibitory effect of lapatinib on T. brucei to HeLa cells. HeLa and T. brucei cells were cultured for 48 h with variable concentrations of lapatinib. Relative cell density represents the live cells expressed as percentages of the control ( i.e. , DMSO-treated) experiment. ( C ) Trypanosome kill assay. Time-course of trypanocidal effect of lapatinib. Trypanosomes (10 6 cells/ml) were treated with DMSO (solvent) or lapatinib (10 µM) for 8 h: viable cells were counted every hour, and plotted for each time point.

    Journal: PLoS ONE

    Article Title: Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds

    doi: 10.1371/journal.pone.0056150

    Figure Lengend Snippet: Lapatinib, AEE788 and canertinib kill bloodstream T. brucei . Bloodstream form T. brucei (initial cell density of 2×10 3 cells/ml) were cultured in 24-well plates for 48 h with either DMSO (control) or different concentrations of drug. Cells were counted with a hemocytometer and the graphs plotted. Data are mean ± standard deviation obtained from two independent experiments performed in duplicate. ( A ). Effect of lapatinib on growth of T. brucei . ( B ). Comparison of growth inhibitory effect of lapatinib on T. brucei to HeLa cells. HeLa and T. brucei cells were cultured for 48 h with variable concentrations of lapatinib. Relative cell density represents the live cells expressed as percentages of the control ( i.e. , DMSO-treated) experiment. ( C ) Trypanosome kill assay. Time-course of trypanocidal effect of lapatinib. Trypanosomes (10 6 cells/ml) were treated with DMSO (solvent) or lapatinib (10 µM) for 8 h: viable cells were counted every hour, and plotted for each time point.

    Article Snippet: AEE788 (Novartis) inhibits EGFR and VEGFR , , and canertinib (CI-1033) (Pfizer) is a pan-inhibitor of EGFRs – .

    Techniques: Cell Culture, Standard Deviation

    Chemical structure of lapatinib, AEE788, and canertinib.

    Journal: PLoS ONE

    Article Title: Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds

    doi: 10.1371/journal.pone.0056150

    Figure Lengend Snippet: Chemical structure of lapatinib, AEE788, and canertinib.

    Article Snippet: AEE788 (Novartis) inhibits EGFR and VEGFR , , and canertinib (CI-1033) (Pfizer) is a pan-inhibitor of EGFRs – .

    Techniques:

    Best models of TbLBPK•drug complexes are consistent with affinity chromatography elution data. ( A ) Predicted ICM binding scores of lapatinib (red), AEE788 (green) and canertinib (magenta) in the binding pockets of the protein kinases. The dotted line represents a hypothetical cutoff for kinase elution by drug after an NAD + wash of the affinity column. ( B ) Predicted binding poses for lapatinib (red), AEE788 (green) and canertinib (magenta) to their highest affinity protein kinases; TbLBPK1, TbLBPK2, and TbCBPK1, respectively. Kinase hinge region is shown in ribbon, ligand atom placement surface is represented by a wire mesh and colored according to its binding properties.

    Journal: PLoS ONE

    Article Title: Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds

    doi: 10.1371/journal.pone.0056150

    Figure Lengend Snippet: Best models of TbLBPK•drug complexes are consistent with affinity chromatography elution data. ( A ) Predicted ICM binding scores of lapatinib (red), AEE788 (green) and canertinib (magenta) in the binding pockets of the protein kinases. The dotted line represents a hypothetical cutoff for kinase elution by drug after an NAD + wash of the affinity column. ( B ) Predicted binding poses for lapatinib (red), AEE788 (green) and canertinib (magenta) to their highest affinity protein kinases; TbLBPK1, TbLBPK2, and TbCBPK1, respectively. Kinase hinge region is shown in ribbon, ligand atom placement surface is represented by a wire mesh and colored according to its binding properties.

    Article Snippet: AEE788 (Novartis) inhibits EGFR and VEGFR , , and canertinib (CI-1033) (Pfizer) is a pan-inhibitor of EGFRs – .

    Techniques: Affinity Chromatography, Binding Assay, Affinity Column

    Dose dependent inhibition of sphere formation by anticancer drugs. To compare two sphere scoring parameters of the SSS vs. “number of spheres”, H1299 (2000 cells per well) were plated in 24-well ULA plates in TS medium supplemented with indicated drugs or vehicle for 4 days. %SSS i and %n i were then calculated. Concentration of drugs were as the following: (A) CI-1033: + 2 μM; (B) Erlotinib: + 3 μM, ++ 6 μM, +++ 10 μM; (C) MK2206: + 0.25 μM, ++ 0.5 μM, +++ 1 μM; (D) Perifosine: + 1 μM, ++ 3 μM, +++ 5 μM; and (E) BEZ235: + 25 nM, ++ 125 nM, +++ 250 nM. The results were presented as Mean ± SEM.

    Journal: PLoS ONE

    Article Title: A Reliable Parameter to Standardize the Scoring of Stem Cell Spheres

    doi: 10.1371/journal.pone.0127348

    Figure Lengend Snippet: Dose dependent inhibition of sphere formation by anticancer drugs. To compare two sphere scoring parameters of the SSS vs. “number of spheres”, H1299 (2000 cells per well) were plated in 24-well ULA plates in TS medium supplemented with indicated drugs or vehicle for 4 days. %SSS i and %n i were then calculated. Concentration of drugs were as the following: (A) CI-1033: + 2 μM; (B) Erlotinib: + 3 μM, ++ 6 μM, +++ 10 μM; (C) MK2206: + 0.25 μM, ++ 0.5 μM, +++ 1 μM; (D) Perifosine: + 1 μM, ++ 3 μM, +++ 5 μM; and (E) BEZ235: + 25 nM, ++ 125 nM, +++ 250 nM. The results were presented as Mean ± SEM.

    Article Snippet: Drugs and treatment Canertinib (CI-1033, Pfizer Pharmaceuticals) and Erlotinib (Selleck Chemicals) were kindly provided by Dr. Mukesh Nyati at University of Michigan.

    Techniques: Inhibition, Concentration Assay

    Growth factors overcome RTK-inhibited-phosphorylation of downstream targets. Examples of immunoblots showing effects of growth factors (100 ng/ml) on Akt and Erk phosphorylation (p) after sorafenib ( A ), dasatinib ( B ), canertinib ( C ), crizotinib ( D ) in pediatric low grade astrocytoma and ependymoma cells (2h). Controls represent the specific cell lines treated with DMSO. Growth-factor-driven rescue on cell viability as given in Fig. 3C and D is indicated underneath the blots: purple squares, no rescue; yellow/orange squares, partial rescue; red squares, complete rescue. Protein expression was normalized to β-actin as loading control and derived from three independent experiments of which the mean (± SEM) was calculated. EGF and HGF-driven rescue resulted in significantly higher protein expression of Akt (p = 0.028) and Erk (p = 0.043) during sorafenib in both UW-467 and Res-196 cells and during dasatinib in the low grade astrocytoma cell lines (p = 0.028 and p = 0.043). Significantly higher Akt phosphorylation was found after HGF addition during canertinib in Res-186, Res-259 and Res-196 cells (p = 0.043) of which Res-186 is shown as an example. *Discrepancies between growth-factor-driven rescue on cell viability level and downstream phosphorylation status.

    Journal: PLoS ONE

    Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma

    doi: 10.1371/journal.pone.0122555

    Figure Lengend Snippet: Growth factors overcome RTK-inhibited-phosphorylation of downstream targets. Examples of immunoblots showing effects of growth factors (100 ng/ml) on Akt and Erk phosphorylation (p) after sorafenib ( A ), dasatinib ( B ), canertinib ( C ), crizotinib ( D ) in pediatric low grade astrocytoma and ependymoma cells (2h). Controls represent the specific cell lines treated with DMSO. Growth-factor-driven rescue on cell viability as given in Fig. 3C and D is indicated underneath the blots: purple squares, no rescue; yellow/orange squares, partial rescue; red squares, complete rescue. Protein expression was normalized to β-actin as loading control and derived from three independent experiments of which the mean (± SEM) was calculated. EGF and HGF-driven rescue resulted in significantly higher protein expression of Akt (p = 0.028) and Erk (p = 0.043) during sorafenib in both UW-467 and Res-196 cells and during dasatinib in the low grade astrocytoma cell lines (p = 0.028 and p = 0.043). Significantly higher Akt phosphorylation was found after HGF addition during canertinib in Res-186, Res-259 and Res-196 cells (p = 0.043) of which Res-186 is shown as an example. *Discrepancies between growth-factor-driven rescue on cell viability level and downstream phosphorylation status.

    Article Snippet: Sorafenib, dasatinib, canertinib and crizotinib (LC laboratories, Woburn MA, USA) were dissolved in sterile dimethyl sulfoxide (DMSO) and stored at -20°C.

    Techniques: Western Blot, Expressing, Derivative Assay

    RTK signaling pathways. Schematic illustration showing important growth factors in brain tumors that can bind RTKs activating downstream pathways which contributes to tumor cell survival. These potential drugable targets can be inhibited by e . g . sorafenib, dasatinib, canertinib and crizotinib.

    Journal: PLoS ONE

    Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma

    doi: 10.1371/journal.pone.0122555

    Figure Lengend Snippet: RTK signaling pathways. Schematic illustration showing important growth factors in brain tumors that can bind RTKs activating downstream pathways which contributes to tumor cell survival. These potential drugable targets can be inhibited by e . g . sorafenib, dasatinib, canertinib and crizotinib.

    Article Snippet: Sorafenib, dasatinib, canertinib and crizotinib (LC laboratories, Woburn MA, USA) were dissolved in sterile dimethyl sulfoxide (DMSO) and stored at -20°C.

    Techniques:

    Combined RTK treatment can overcome growth-factor-driven rescue. Cell viability assays demonstrating additional effects of combination treatment in pediatric low grade astrocytoma (Res-259) and ependymoma (Res-196) compared with single treatment (mean ± SD, 48h). Immunoblots showing effect of single RTK inhibition (8 uM), the rescue effect of growth factors (100 ng/ml) and the ability of the second inhibitor (canertinib 4 uM, crizotinib 3 uM, sorafenib 2 uM) to overcome this growth-factor-driven rescue on Akt and Erk phosphorylation (p) in pediatric low grade astrocytoma and ependymoma cells (2h). Protein expression was normalized to β-actin as loading control and derived from three independent experiments of which the mean (± SEM) was calculated. The ability of crizotinib to overcome HGF-driven rescue during canertinib treatment reached statistical significance on Akt and Erk phosphorylation (p = 0.028 and p = 0.046 respectively).

    Journal: PLoS ONE

    Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma

    doi: 10.1371/journal.pone.0122555

    Figure Lengend Snippet: Combined RTK treatment can overcome growth-factor-driven rescue. Cell viability assays demonstrating additional effects of combination treatment in pediatric low grade astrocytoma (Res-259) and ependymoma (Res-196) compared with single treatment (mean ± SD, 48h). Immunoblots showing effect of single RTK inhibition (8 uM), the rescue effect of growth factors (100 ng/ml) and the ability of the second inhibitor (canertinib 4 uM, crizotinib 3 uM, sorafenib 2 uM) to overcome this growth-factor-driven rescue on Akt and Erk phosphorylation (p) in pediatric low grade astrocytoma and ependymoma cells (2h). Protein expression was normalized to β-actin as loading control and derived from three independent experiments of which the mean (± SEM) was calculated. The ability of crizotinib to overcome HGF-driven rescue during canertinib treatment reached statistical significance on Akt and Erk phosphorylation (p = 0.028 and p = 0.046 respectively).

    Article Snippet: Sorafenib, dasatinib, canertinib and crizotinib (LC laboratories, Woburn MA, USA) were dissolved in sterile dimethyl sulfoxide (DMSO) and stored at -20°C.

    Techniques: Western Blot, Inhibition, Expressing, Derivative Assay

    Tumor cell migration. Cell monolayers were scratched with a pipette tip to produce a clear cell-free area into which cells at the periphery could migrate. Micrographs showing migrated Res-259 cells treated with DMSO (control), the effect on tumor cell migration of HGF (100 ng/ml), canertinib (2 uM), HGF addition to canertinib and the combination treatment including canertinib and crizotinib (3 uM) after 24h (upper panels) and 48h (lower panels).

    Journal: PLoS ONE

    Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma

    doi: 10.1371/journal.pone.0122555

    Figure Lengend Snippet: Tumor cell migration. Cell monolayers were scratched with a pipette tip to produce a clear cell-free area into which cells at the periphery could migrate. Micrographs showing migrated Res-259 cells treated with DMSO (control), the effect on tumor cell migration of HGF (100 ng/ml), canertinib (2 uM), HGF addition to canertinib and the combination treatment including canertinib and crizotinib (3 uM) after 24h (upper panels) and 48h (lower panels).

    Article Snippet: Sorafenib, dasatinib, canertinib and crizotinib (LC laboratories, Woburn MA, USA) were dissolved in sterile dimethyl sulfoxide (DMSO) and stored at -20°C.

    Techniques: Migration, Transferring

    RTK expression and inhibition. A , Heat map showing percentages of cells expressing numerous growth factor receptors above control levels using flow cytometry analyses in pediatric low grade astrocytoma (Res-186, Res-259, UW-467) and ependymoma (Res-196) cell lines. B , Cell viability assays demonstrating the mean effect ± SD of sorafenib, dasatinib, canertinib and crizotinib on pediatric low grade astrocytoma (Res-186, Res-259, UW-467) and ependymoma (Res-196) cell lines after 48h.

    Journal: PLoS ONE

    Article Title: Growth-Factor-Driven Rescue to Receptor Tyrosine Kinase (RTK) Inhibitors through Akt and Erk Phosphorylation in Pediatric Low Grade Astrocytoma and Ependymoma

    doi: 10.1371/journal.pone.0122555

    Figure Lengend Snippet: RTK expression and inhibition. A , Heat map showing percentages of cells expressing numerous growth factor receptors above control levels using flow cytometry analyses in pediatric low grade astrocytoma (Res-186, Res-259, UW-467) and ependymoma (Res-196) cell lines. B , Cell viability assays demonstrating the mean effect ± SD of sorafenib, dasatinib, canertinib and crizotinib on pediatric low grade astrocytoma (Res-186, Res-259, UW-467) and ependymoma (Res-196) cell lines after 48h.

    Article Snippet: Sorafenib, dasatinib, canertinib and crizotinib (LC laboratories, Woburn MA, USA) were dissolved in sterile dimethyl sulfoxide (DMSO) and stored at -20°C.

    Techniques: Expressing, Inhibition, Flow Cytometry, Cytometry

    HPV8E6 directly binds to the ubiquitinated EGFR in an UV-dependent manner. (A) RTS3b cells were transiently transfected with a vector directing the expression of FLAG-tagged HPV8E6 under control of the CMV-promoter or the empty vector. Where indicated the cells have been irradiated with UV-light 30 min prior harvesting. 400 μg of extracts were incubated with the anti-EGFR-antibody (lanes 1–4) or with the FLAG-antibody (lanes 5–8), both coupled to sepharose, washed four times in 0.3 M KCl-buffer prior analysis by WB and 40 μg of the extracts were used in the input blots. (B) IP with anti-EGFR sepharose and 800 μg of extracts derived from the stable RTS3b-pLXSN and RTS3b-HPV8E6 lines, either left untreated or UV-irradiated 30 min prior harvesting. FLAG-HPV8E6 and EGFR present in the precipitate as well as in the input were detected by WB. (C) Extracts from UV-irradiated RTS3b cells that have been transiently transfected with the FLAG-HPV8E6 or the FLAG-vector were incubated either with DMSO (lanes 1, 2, 5), 3 μM (lane 3) or 7 μM (lane 4) Canertinib for 30 min. The IP was performed with the FLAG-antibody. All WB were developed with the indicated antibodies. In lanes 6–10, C33A cells were transiently transfected with expression vectors for EGFR-GFP and FLAG-HPV8E6, and UV-irradiated, where indicated, prior harvesting 30 min later. EGFR-GFP was precipitated from 400 μg of extract by the EGFR antibody and bound HPV8E6 was detected by the FLAG-antibody. The input blots were developed with the antibodies against pEGFR-Y1068, GFP, and FLAG. The positions of the molecular weight markers are provided (in kDa).

    Journal: Frontiers in Microbiology

    Article Title: Induction of Tyrosine Phosphorylation of UV-Activated EGFR by the Beta-Human Papillomavirus Type 8 E6 Leads to Papillomatosis

    doi: 10.3389/fmicb.2017.02197

    Figure Lengend Snippet: HPV8E6 directly binds to the ubiquitinated EGFR in an UV-dependent manner. (A) RTS3b cells were transiently transfected with a vector directing the expression of FLAG-tagged HPV8E6 under control of the CMV-promoter or the empty vector. Where indicated the cells have been irradiated with UV-light 30 min prior harvesting. 400 μg of extracts were incubated with the anti-EGFR-antibody (lanes 1–4) or with the FLAG-antibody (lanes 5–8), both coupled to sepharose, washed four times in 0.3 M KCl-buffer prior analysis by WB and 40 μg of the extracts were used in the input blots. (B) IP with anti-EGFR sepharose and 800 μg of extracts derived from the stable RTS3b-pLXSN and RTS3b-HPV8E6 lines, either left untreated or UV-irradiated 30 min prior harvesting. FLAG-HPV8E6 and EGFR present in the precipitate as well as in the input were detected by WB. (C) Extracts from UV-irradiated RTS3b cells that have been transiently transfected with the FLAG-HPV8E6 or the FLAG-vector were incubated either with DMSO (lanes 1, 2, 5), 3 μM (lane 3) or 7 μM (lane 4) Canertinib for 30 min. The IP was performed with the FLAG-antibody. All WB were developed with the indicated antibodies. In lanes 6–10, C33A cells were transiently transfected with expression vectors for EGFR-GFP and FLAG-HPV8E6, and UV-irradiated, where indicated, prior harvesting 30 min later. EGFR-GFP was precipitated from 400 μg of extract by the EGFR antibody and bound HPV8E6 was detected by the FLAG-antibody. The input blots were developed with the antibodies against pEGFR-Y1068, GFP, and FLAG. The positions of the molecular weight markers are provided (in kDa).

    Article Snippet: The Canertinib group was administered once 20 mg/kg Canertinib (CI-1033, Selleckchem, S1019) solved in 150 μl PBS by gavage and the control group obtained 150 μl PBS 4 h prior UV-irradiation.

    Techniques: Transfection, Plasmid Preparation, Expressing, Irradiation, Incubation, Western Blot, Derivative Assay, Molecular Weight

    Inhibition of the RTK-activity of the EGFR by the pan-ErbB-inhibitor Canertinib (CI-1033) suppresses UV-induced papillomatosis in K14-HPV8E6 transgenic mice. (A) Immunohistochemical staining of paraffin embedded sections obtained from skin of K14-HPV8E6 transgenic mice 13 days after UV-exposure with the anti-pEGFR-Y1068 or rabbit IgG isotype control (magnification 1:400). (B) Transgenic mice obtained either Canertinib (CI-1033) (20 mg/kg weight) or the same volume of PBS 4 h prior UV-irradiation followed by applications of Canertinib once daily at a dose of 10 mg/kg weight or 150 μl PBS for 24 days, as given in the time line. The pictures represent examples of the lesions from four mice treated with CI-1033 (M19, 20, 23, and M26) and from one control mouse which obtained PBS (M22). The graph beneath shows the quantification of RT-PCR results with RNA isolated from two PBS and three Canertinib-treated animals to demonstrate the expression of HPV8E6. (C) Histology with H/E staining of skin section obtained from two PBS-control (M21 and M22) and four Canertinib (CI-1033)-treated animals (M19, M20, M23, and M26) 24 days after UV-irradiation.

    Journal: Frontiers in Microbiology

    Article Title: Induction of Tyrosine Phosphorylation of UV-Activated EGFR by the Beta-Human Papillomavirus Type 8 E6 Leads to Papillomatosis

    doi: 10.3389/fmicb.2017.02197

    Figure Lengend Snippet: Inhibition of the RTK-activity of the EGFR by the pan-ErbB-inhibitor Canertinib (CI-1033) suppresses UV-induced papillomatosis in K14-HPV8E6 transgenic mice. (A) Immunohistochemical staining of paraffin embedded sections obtained from skin of K14-HPV8E6 transgenic mice 13 days after UV-exposure with the anti-pEGFR-Y1068 or rabbit IgG isotype control (magnification 1:400). (B) Transgenic mice obtained either Canertinib (CI-1033) (20 mg/kg weight) or the same volume of PBS 4 h prior UV-irradiation followed by applications of Canertinib once daily at a dose of 10 mg/kg weight or 150 μl PBS for 24 days, as given in the time line. The pictures represent examples of the lesions from four mice treated with CI-1033 (M19, 20, 23, and M26) and from one control mouse which obtained PBS (M22). The graph beneath shows the quantification of RT-PCR results with RNA isolated from two PBS and three Canertinib-treated animals to demonstrate the expression of HPV8E6. (C) Histology with H/E staining of skin section obtained from two PBS-control (M21 and M22) and four Canertinib (CI-1033)-treated animals (M19, M20, M23, and M26) 24 days after UV-irradiation.

    Article Snippet: The Canertinib group was administered once 20 mg/kg Canertinib (CI-1033, Selleckchem, S1019) solved in 150 μl PBS by gavage and the control group obtained 150 μl PBS 4 h prior UV-irradiation.

    Techniques: Inhibition, Activity Assay, Transgenic Assay, Mouse Assay, Immunohistochemistry, Staining, Irradiation, Reverse Transcription Polymerase Chain Reaction, Isolation, Expressing

    Selection of canertinib resistant cells. Tu-2449 cells were treated with canertinib for 10 days. The drug was replenished in the growth medium in 24 hour intervals by half-medium change. After 10 days, drug treatment was discontinued for three days. Cell growth was microscopically monitored after 1, 3, 5, 7, 9 and 13 days. The upper panel shows different fields while the lower panel shows the same field. Magnification, 40×. Drug treatment selects for small cell colonies with higher cell density.

    Journal: MedChemComm

    Article Title: Tumor cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib †The authors declare no conflict of interest.

    doi: 10.1039/c6md00463f

    Figure Lengend Snippet: Selection of canertinib resistant cells. Tu-2449 cells were treated with canertinib for 10 days. The drug was replenished in the growth medium in 24 hour intervals by half-medium change. After 10 days, drug treatment was discontinued for three days. Cell growth was microscopically monitored after 1, 3, 5, 7, 9 and 13 days. The upper panel shows different fields while the lower panel shows the same field. Magnification, 40×. Drug treatment selects for small cell colonies with higher cell density.

    Article Snippet: Chemicals The tyrosine kinase inhibitor canertinib (CI-1033) was purchased from SelleckChem, Munich, Germany, dissolved in sterile dimethyl sulfoxide (DMSO) and stored at –80 °C.

    Techniques: Selection

    Selected canertinib resistant colonies become sensitive again to drug treatment when cultured at low cell densities. Parental and 5 selected colonies of Tu-2449 glioma cells were plated at low densities (comparable to Fig. 2 panel one) and treated with 5 μM canertinib or only the growth medium. The drug was replenished in the growth medium in 24 hour intervals. Cell viability was monitored after 24, 48 and 72 h using the AlamarBlue® assay. Repeated addition of 5 μM canertinib significantly reduced the cell viability in all analyzed cell clones after 72 h. Results are presented as mean ± SD, with respect to 72 h untreated cells; **** p

    Journal: MedChemComm

    Article Title: Tumor cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib †The authors declare no conflict of interest.

    doi: 10.1039/c6md00463f

    Figure Lengend Snippet: Selected canertinib resistant colonies become sensitive again to drug treatment when cultured at low cell densities. Parental and 5 selected colonies of Tu-2449 glioma cells were plated at low densities (comparable to Fig. 2 panel one) and treated with 5 μM canertinib or only the growth medium. The drug was replenished in the growth medium in 24 hour intervals. Cell viability was monitored after 24, 48 and 72 h using the AlamarBlue® assay. Repeated addition of 5 μM canertinib significantly reduced the cell viability in all analyzed cell clones after 72 h. Results are presented as mean ± SD, with respect to 72 h untreated cells; **** p

    Article Snippet: Chemicals The tyrosine kinase inhibitor canertinib (CI-1033) was purchased from SelleckChem, Munich, Germany, dissolved in sterile dimethyl sulfoxide (DMSO) and stored at –80 °C.

    Techniques: Cell Culture, Alamar Blue Assay, Clone Assay

    Canertinib treatment of Tu-2449 glioma cells induces the arrest of cell growth after 24 h and prolonged inhibition causes cell death dependent upon exposure time and drug dose. (A) Tu-2449 cells were treated with a single dose of 5 or 10 μM canertinib or DMSO and cultured for 24 h. Microscopic monitoring after 24 h reveals growth arrest in canertinib treated cells; scale bar: 100 μm. (B) Tu-2449 cells were treated with canertinib or DMSO and the drug was replenished in the growth medium in 24 hour intervals. Cell viability was monitored after 24, 48 and 72 h using the AlamarBlue® assay. Repeated addition of canertinib arrests the cells and causes cell death after 24 h (10 μM canertinib) or 48 h (5 μM canertinib). Results are presented as mean ± SD, expressed as a percentage with respect to 72 h DMSO. ns = not significant, **** p

    Journal: MedChemComm

    Article Title: Tumor cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib †The authors declare no conflict of interest.

    doi: 10.1039/c6md00463f

    Figure Lengend Snippet: Canertinib treatment of Tu-2449 glioma cells induces the arrest of cell growth after 24 h and prolonged inhibition causes cell death dependent upon exposure time and drug dose. (A) Tu-2449 cells were treated with a single dose of 5 or 10 μM canertinib or DMSO and cultured for 24 h. Microscopic monitoring after 24 h reveals growth arrest in canertinib treated cells; scale bar: 100 μm. (B) Tu-2449 cells were treated with canertinib or DMSO and the drug was replenished in the growth medium in 24 hour intervals. Cell viability was monitored after 24, 48 and 72 h using the AlamarBlue® assay. Repeated addition of canertinib arrests the cells and causes cell death after 24 h (10 μM canertinib) or 48 h (5 μM canertinib). Results are presented as mean ± SD, expressed as a percentage with respect to 72 h DMSO. ns = not significant, **** p

    Article Snippet: Chemicals The tyrosine kinase inhibitor canertinib (CI-1033) was purchased from SelleckChem, Munich, Germany, dissolved in sterile dimethyl sulfoxide (DMSO) and stored at –80 °C.

    Techniques: Inhibition, Cell Culture, Alamar Blue Assay

    APAP caused rapid mitochondrial translocation of EGFR and mitochondrial dysfunction, which was inhibited by EGFRi. A, Western blot analysis of phospho-EGFR and EGFR in mitochondrial fraction at 1.5 h after administration of APAP (300 mg/kg) in mice pretreated (2-h prior to APAP) with canertinib (80 mg/kg). B-E , Oxygen consumption rate analysis in freshly isolated mitochondria from mice ( n = 3) treated with APAP (300 mg/kg) + PBS, APAP (300 mg/kg) + canertinib (80 mg/kg) or saline (control), using Seahorse extracellular flux analyzer. B and C, Canertinib was administered 2hr before APAP and analysis was done 1.5 h after APAP treatment. D and E, Canertinib was administered 1-h post-APAP and analysis was done 3 h after APAP treatment. Oxidative phosphorylation was manipulated with injection of ADP, oligomycin (oligo), FCCP, and antimycin A (anti-A). Respiration was sequentially measured in a coupled state with substrate present (basal respiration), followed by State 3 (phosphorylating respiration, in the presence of ADP and substrate), State 4o (leak state induced with the addition of oligomycin - inhibitor of ATP synthase), and then maximal uncoupler (FCCP)-stimulated respiration (State 3u). At the end, antimycin A (complex III inhibitor) was added to inhibit mitochondrial respiration completely * indicate significant difference w.r.t. to APAP + PBS group at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: APAP caused rapid mitochondrial translocation of EGFR and mitochondrial dysfunction, which was inhibited by EGFRi. A, Western blot analysis of phospho-EGFR and EGFR in mitochondrial fraction at 1.5 h after administration of APAP (300 mg/kg) in mice pretreated (2-h prior to APAP) with canertinib (80 mg/kg). B-E , Oxygen consumption rate analysis in freshly isolated mitochondria from mice ( n = 3) treated with APAP (300 mg/kg) + PBS, APAP (300 mg/kg) + canertinib (80 mg/kg) or saline (control), using Seahorse extracellular flux analyzer. B and C, Canertinib was administered 2hr before APAP and analysis was done 1.5 h after APAP treatment. D and E, Canertinib was administered 1-h post-APAP and analysis was done 3 h after APAP treatment. Oxidative phosphorylation was manipulated with injection of ADP, oligomycin (oligo), FCCP, and antimycin A (anti-A). Respiration was sequentially measured in a coupled state with substrate present (basal respiration), followed by State 3 (phosphorylating respiration, in the presence of ADP and substrate), State 4o (leak state induced with the addition of oligomycin - inhibitor of ATP synthase), and then maximal uncoupler (FCCP)-stimulated respiration (State 3u). At the end, antimycin A (complex III inhibitor) was added to inhibit mitochondrial respiration completely * indicate significant difference w.r.t. to APAP + PBS group at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Translocation Assay, Western Blot, Mouse Assay, Isolation, Injection

    Decreased oxidative stress, mitochondrial protein nitration and release of endonucleases from mitochondria by EGFRi. A, Oxidized glutathione levels in total liver extract. B, Representative photomicrographs of liver sections stained for nitrotyrosine-protein adducts. C, Western blot analysis of nitrotyrosine adducts in mitochondrial fraction with its densitometric analysis shown in (D) . E, Western blot analysis of AIF, endonuclease G and SMAC in cytosolic fraction with densitometric analysis of AIF as shown in (F) . All analysis were done on liver samples collected at various time points after treatment with APAP (300 mg/kg) + PBS or APAP (300 mg/kg) + canertinib (80 mg/kg, 1-h post-APAP) ( n = 3–5). *indicate significant difference between groups at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: Decreased oxidative stress, mitochondrial protein nitration and release of endonucleases from mitochondria by EGFRi. A, Oxidized glutathione levels in total liver extract. B, Representative photomicrographs of liver sections stained for nitrotyrosine-protein adducts. C, Western blot analysis of nitrotyrosine adducts in mitochondrial fraction with its densitometric analysis shown in (D) . E, Western blot analysis of AIF, endonuclease G and SMAC in cytosolic fraction with densitometric analysis of AIF as shown in (F) . All analysis were done on liver samples collected at various time points after treatment with APAP (300 mg/kg) + PBS or APAP (300 mg/kg) + canertinib (80 mg/kg, 1-h post-APAP) ( n = 3–5). *indicate significant difference between groups at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Nitration, Staining, Western Blot

    Impaired liver regeneration and cell cycle arrest by delayed (12-h post-APAP) treatment with EGFRi. A , Representative photomicrographs of PCNA stained liver sections with arrows indicating cells in S-phase with nuclear PCNA staining (brown). B, Total number of PCNA-positive cell per high power field (×40). C, Western blot analysis of cyclinD1, CDK4, phospho-Rb, and PCNA using total liver extract. Densitometric analysis of (D) cyclinD1, (E) CDK4, and (F) PCNA. Mice were treated with 300 mg/kg APAP. Canertinib (80 mg/kg) or PBS was administered 12-h post-APAP ( n = 3–5). All samples were collected at 24 and 48 h after APAP treatment. * indicate significant difference between groups at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: Impaired liver regeneration and cell cycle arrest by delayed (12-h post-APAP) treatment with EGFRi. A , Representative photomicrographs of PCNA stained liver sections with arrows indicating cells in S-phase with nuclear PCNA staining (brown). B, Total number of PCNA-positive cell per high power field (×40). C, Western blot analysis of cyclinD1, CDK4, phospho-Rb, and PCNA using total liver extract. Densitometric analysis of (D) cyclinD1, (E) CDK4, and (F) PCNA. Mice were treated with 300 mg/kg APAP. Canertinib (80 mg/kg) or PBS was administered 12-h post-APAP ( n = 3–5). All samples were collected at 24 and 48 h after APAP treatment. * indicate significant difference between groups at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Staining, Western Blot, Mouse Assay

    Increased progression of injury, impaired recovery and decreased survival after delayed (12 hr post-APAP) treatment with EGFRi. (A) Schematic showing experimental design. (B) Representative photomicrographs of H E stained liver sections with necrotic area outlined, (C) serum ALT levels with percentage survival specified over bars and (D) percentage necrosis area based on H E stained liver sections of mice treated with 300 mg/kg APAP followed by treatment with canertinib (80 mg/kg) or PBS, 12 hr post-APAP. All samples were collected and survival was recorded at 24 and 48 hr after APAP treatment (n=5-6). * indicate significant difference between groups at p

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: Increased progression of injury, impaired recovery and decreased survival after delayed (12 hr post-APAP) treatment with EGFRi. (A) Schematic showing experimental design. (B) Representative photomicrographs of H E stained liver sections with necrotic area outlined, (C) serum ALT levels with percentage survival specified over bars and (D) percentage necrosis area based on H E stained liver sections of mice treated with 300 mg/kg APAP followed by treatment with canertinib (80 mg/kg) or PBS, 12 hr post-APAP. All samples were collected and survival was recorded at 24 and 48 hr after APAP treatment (n=5-6). * indicate significant difference between groups at p

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Staining, Mouse Assay

    Early treatment with EGFRi (1-h post-APAP) remarkably attenuated APAP-induced hepatotoxicity without altering APAP bioactivation and APAP-protein adducts formation. A, Western blot analysis of phospho-EGFR and EGFR in liver lysate, B, representative photomicrographs of H E stained liver sections with necrotic area outlined, C, serum ALT levels, D, percentage necrosis area based on H E stained liver sections, E, total glutathione levels in liver extract, and F, APAP-protein adducts levels in liver as measured by HPLC-ECD method. For all experiments mice ( n = 3–12) were treated with 300 mg/kg APAP followed by canertinib (80 mg/kg) or PBS, 1-h post-APAP. All samples were collected at various time points up-to 24-h after APAP treatment. * indicate significant difference between groups at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: Early treatment with EGFRi (1-h post-APAP) remarkably attenuated APAP-induced hepatotoxicity without altering APAP bioactivation and APAP-protein adducts formation. A, Western blot analysis of phospho-EGFR and EGFR in liver lysate, B, representative photomicrographs of H E stained liver sections with necrotic area outlined, C, serum ALT levels, D, percentage necrosis area based on H E stained liver sections, E, total glutathione levels in liver extract, and F, APAP-protein adducts levels in liver as measured by HPLC-ECD method. For all experiments mice ( n = 3–12) were treated with 300 mg/kg APAP followed by canertinib (80 mg/kg) or PBS, 1-h post-APAP. All samples were collected at various time points up-to 24-h after APAP treatment. * indicate significant difference between groups at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Western Blot, Staining, High Performance Liquid Chromatography, Mouse Assay

    JNK activation, its mitochondrial translocation and signaling through other protein kinases not altered by EGFRi. Western blot analysis of phospho-JNK and JNK in (A) total cell lysate and (B) mitochondrial fraction with densitometric analysis of mitochondrial activated JNK (p-JNK) shown in (C) . D, Western blot analysis of RIP3 and RIP1 in total cell lysate. E, Western blot analysis showing PKC activation (studied using antibody against phosphorylated PKC-substrates in mitochondria) with its densitometric analysis shown in (F) . All analysis were done on liver samples collected at various time points after treatment with 300 mg/kg APAP + PBS or APAP + canertinib (80 mg/kg, 1-h post-APAP) ( n = 3–4). *indicate significant difference between groups at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: JNK activation, its mitochondrial translocation and signaling through other protein kinases not altered by EGFRi. Western blot analysis of phospho-JNK and JNK in (A) total cell lysate and (B) mitochondrial fraction with densitometric analysis of mitochondrial activated JNK (p-JNK) shown in (C) . D, Western blot analysis of RIP3 and RIP1 in total cell lysate. E, Western blot analysis showing PKC activation (studied using antibody against phosphorylated PKC-substrates in mitochondria) with its densitometric analysis shown in (F) . All analysis were done on liver samples collected at various time points after treatment with 300 mg/kg APAP + PBS or APAP + canertinib (80 mg/kg, 1-h post-APAP) ( n = 3–4). *indicate significant difference between groups at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Activation Assay, Translocation Assay, Western Blot

    (A–C) Role of glutathione depletion in rapid activation of EGFR by APAP. (D–F) Enhanced protection against APAP hepatotoxicity by combination of NAC and EGFRi (administered 4-h post-APAP). (A) Total glutathione levels in liver extract and (B) western blot analysis of phospho-EGFR and EGFR in total cell lysate from liver samples obtained 2 h after treatment with Phorone (200 mg/kg in corn oil) or corn oil (control) in mice ( n = 5). (C) Densitomertic analysis showing EGFR activation based on western blot image shown in (B) . (D) Representative photomicrographs of H E stained liver sections with necrotic area outlined, (E) serum ALT levels and (F) percentage necrosis area based on H E stained liver sections of mice treated with 300 mg/kg APAP followed by canertinib (80 mg/kg), NAC (500 mg/kg), combination of canertinib (80 mg/kg) and NAC (500 mg/kg) or PBS (control), 4-h post-APAP ( n = 5). Samples were collected 24 h after APAP treatment. *indicate significant difference w.r.t. control group at P

    Journal: Toxicological Sciences

    Article Title: Dual Role of Epidermal Growth Factor Receptor in Liver Injury and Regeneration after Acetaminophen Overdose in Mice

    doi: 10.1093/toxsci/kfw213

    Figure Lengend Snippet: (A–C) Role of glutathione depletion in rapid activation of EGFR by APAP. (D–F) Enhanced protection against APAP hepatotoxicity by combination of NAC and EGFRi (administered 4-h post-APAP). (A) Total glutathione levels in liver extract and (B) western blot analysis of phospho-EGFR and EGFR in total cell lysate from liver samples obtained 2 h after treatment with Phorone (200 mg/kg in corn oil) or corn oil (control) in mice ( n = 5). (C) Densitomertic analysis showing EGFR activation based on western blot image shown in (B) . (D) Representative photomicrographs of H E stained liver sections with necrotic area outlined, (E) serum ALT levels and (F) percentage necrosis area based on H E stained liver sections of mice treated with 300 mg/kg APAP followed by canertinib (80 mg/kg), NAC (500 mg/kg), combination of canertinib (80 mg/kg) and NAC (500 mg/kg) or PBS (control), 4-h post-APAP ( n = 5). Samples were collected 24 h after APAP treatment. *indicate significant difference w.r.t. control group at P

    Article Snippet: For studies with EGFR inhibitor (EGFRi), canertinib dihydrochloride salt (80 mg/kg, i.p.) (LC Laboratories, Woburn, Massachusetts) was dissolved in warm phosphate-buffered saline (PBS).

    Techniques: Activation Assay, Western Blot, Mouse Assay, Staining

    Modulation of PI3-K activity inversely impacts InlB-requirement for Lm entry in vitro. (A) Whole cell extracts from LS174T cells incubated with PBS or different concentrations of the ErbB inhibitor CI-1033 (0.01, 0.1, or 1 µM) during 24 h, immunoblotted for P-Akt and total Akt. (B) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in LS174T cells incubated with PBS (controls, black bars) or CI-1033 1 µM (gray bars) for 24 h. (C) Whole cell extracts from LS174T cells incubated with PBS or CI-1033 1 µM and infected with WT Lm EGD and isogenic mutant Δ inlB , immunoblotted for P-Akt and total Akt. (D) Whole cell extracts from Jar cells incubated with PBS or EGF 50 ng/ml during 10, 30, or 60 min, immunoblotted for P-Akt and total Akt. (E) Whole cell extracts from Jar cells transfected with esiRNA PTEN, immunoblotted for PTEN. (F) Whole cell extracts from Jar cells transfected with esiRNA PTEN or control scrambled and incubated with PBS or EGF 50 ng/ml for 120 min, immunoblotted for P-Akt and total Akt. (G) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS, or transfected with esiRNA PTEN or incubated with EGF 50 ng/ml. (H) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS or incubated with EGF 50 ng/ml, and transfected with esiRNA PTEN. (B, G, and H) Invasion assays were performed in triplicates and represent at least three independent experiments for each condition tested. Relative entry represents the ratio of CFUs for each strain divided by the mean of CFUs for the WT Lm EGD strain. A one-way ANOVA test followed by a Bonferroni’s multiple comparisons was performed. Error bars, SEM. (A, C, D, E, and F) Immunoblots were performed at least two times. Densitometry was performed and the ratio of P-Akt over total Akt is relative to the PBS, for which the value is normalized to 1.

    Journal: The Journal of Experimental Medicine

    Article Title: PI3-kinase activation is critical for host barrier permissiveness to Listeria monocytogenes

    doi: 10.1084/jem.20141406

    Figure Lengend Snippet: Modulation of PI3-K activity inversely impacts InlB-requirement for Lm entry in vitro. (A) Whole cell extracts from LS174T cells incubated with PBS or different concentrations of the ErbB inhibitor CI-1033 (0.01, 0.1, or 1 µM) during 24 h, immunoblotted for P-Akt and total Akt. (B) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in LS174T cells incubated with PBS (controls, black bars) or CI-1033 1 µM (gray bars) for 24 h. (C) Whole cell extracts from LS174T cells incubated with PBS or CI-1033 1 µM and infected with WT Lm EGD and isogenic mutant Δ inlB , immunoblotted for P-Akt and total Akt. (D) Whole cell extracts from Jar cells incubated with PBS or EGF 50 ng/ml during 10, 30, or 60 min, immunoblotted for P-Akt and total Akt. (E) Whole cell extracts from Jar cells transfected with esiRNA PTEN, immunoblotted for PTEN. (F) Whole cell extracts from Jar cells transfected with esiRNA PTEN or control scrambled and incubated with PBS or EGF 50 ng/ml for 120 min, immunoblotted for P-Akt and total Akt. (G) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS, or transfected with esiRNA PTEN or incubated with EGF 50 ng/ml. (H) Invasion assays of WT Lm EGD and isogenic mutants Δ inlA and Δ inlB in Jar cells incubated with PBS or incubated with EGF 50 ng/ml, and transfected with esiRNA PTEN. (B, G, and H) Invasion assays were performed in triplicates and represent at least three independent experiments for each condition tested. Relative entry represents the ratio of CFUs for each strain divided by the mean of CFUs for the WT Lm EGD strain. A one-way ANOVA test followed by a Bonferroni’s multiple comparisons was performed. Error bars, SEM. (A, C, D, E, and F) Immunoblots were performed at least two times. Densitometry was performed and the ratio of P-Akt over total Akt is relative to the PBS, for which the value is normalized to 1.

    Article Snippet: The pan-ErbB inhibitor CI-1033 was purchased from Biovision.

    Techniques: Activity Assay, In Vitro, Incubation, Infection, Mutagenesis, Transfection, esiRNA, Western Blot

    Validation of selected GenEx-TNBC predictions in TNBC cell lines of multiple molecular subtypes Crystal violet staining of HCC1937 (BL1), BT549 (ML), and MDA-MB-453 (LAR) cells grown in the presence of the indicated concentrations of Mecamylamine (A) , Canertinib (B) , Bortezomib (C) , and Tretinoin (D) for seven (7) days. Data for each cell line were normalized to the appropriate solvent control (ethanol for Mecamylamine, DMSO for all others). Data were analyzed by two-way ANOVA with Tukey post hoc multiple comparisons test (asterisks indicate per-dose comparisons) and are presented as the mean +/− standard deviation (S.D.) for a single experiment performed with 6 technical replicates that is representative of at least two independent experiments (biological replicates).

    Journal: Oncotarget

    Article Title: Differential prioritization of therapies to subtypes of triple negative breast cancer using a systems medicine method

    doi: 10.18632/oncotarget.21669

    Figure Lengend Snippet: Validation of selected GenEx-TNBC predictions in TNBC cell lines of multiple molecular subtypes Crystal violet staining of HCC1937 (BL1), BT549 (ML), and MDA-MB-453 (LAR) cells grown in the presence of the indicated concentrations of Mecamylamine (A) , Canertinib (B) , Bortezomib (C) , and Tretinoin (D) for seven (7) days. Data for each cell line were normalized to the appropriate solvent control (ethanol for Mecamylamine, DMSO for all others). Data were analyzed by two-way ANOVA with Tukey post hoc multiple comparisons test (asterisks indicate per-dose comparisons) and are presented as the mean +/− standard deviation (S.D.) for a single experiment performed with 6 technical replicates that is representative of at least two independent experiments (biological replicates).

    Article Snippet: Compounds Mecamylamine (Sigma Aldrich), Canertinib (AK Scientific, Union City, CA), Bortezomib (AK Scientific), Tretinoin (AK Scientific), and AMG900 (Selleckchem, Houston, TX) were resuspended in dimethyl sulfoxide (DMSO – Canertinib, Bortezomib, Tretinoin, AMG900) or 200 proof ethanol (EtOH – Mecamylamine) at a concentration of 10 mM, stored at -20°C, and used at the indicated concentrations.

    Techniques: Staining, Multiple Displacement Amplification, Standard Deviation