rabbit polyclonal antibodies against egfr  (Cell Signaling Technology Inc)

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Antitumor activity of afatinib in EGFR T790M-negative human oral cancer therapeutically targets mTOR/Mcl-1 signaling axis

doi: 10.1007/s13402-024-00962-6

Figure Lengend Snippet: The correlation between EGFR and Mcl-1 expressions in HNC and oral cancer patients. A . Analysis of the correlation between EGFR and Mcl-1 mRNA expressions in HNC patients from the TCGA dataset. B . KM plotter analysis of the association between concurrent expression of EGFR and Mcl-1 and survival rate in HNC patients. C . IHC analysis of the association between the protein expression levels of p-EGFR and Mcl-1 in the tissues of 52 oral cancer patients ( upper panel ), The representative images of p-EGFR and Mcl-1 expression (cases 49, 39, and 34 for pEGFR+/Mcl-1 + and case 44 for pEGFR-/Mcl-1-) ( lower panel ) the graphical pattern of the corresponding IHC scores. D . Selection of oral cancer cell lines with high coexpression of p-EGFR and Mcl-1 (indicated by red box) using Western blotting in HOK and 13 oral cancer cell lines

Article Snippet: The following primary antibodies were used in the experiments: rabbit antihuman polyclonal antibodies against cleaved PARP (Cat. No. 9541; 1:1000), cleaved caspase-3 (Cat. No. 9664; 1:1000), phospho-mTOR (Cat. No. 2971; 1:1000), phospho-p70S6 (Cat. No. 9205; 1:1000), Bim (Cat. No. 2819; 1:1000), Mcl-1 (Cat. No. 4572; 1:1000), and rabbit antihuman monoclonal antibodies against phospho-EGFR (Cat. No. 3777; 1:1000), mTOR (Cat. No. 2983; 1:1000), and c-MYC (Cat. No. 5605; 1:1000) that were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA); and rabbit antihuman polyclonal antibody against EGFR (Cat. No. sc-03; 1:1000), goat antihuman polyclonal antibody against t-Bid (Cat. No. 34,325; 1:1000), and mouse antihuman monoclonal antibodies against β-actin (Cat. No. 47,778; 1:3000) and GAPDH (Cat. No. ab9484; 1:3000) that were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA).

Techniques: Expressing, Selection, Western Blot

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Antitumor activity of afatinib in EGFR T790M-negative human oral cancer therapeutically targets mTOR/Mcl-1 signaling axis

doi: 10.1007/s13402-024-00962-6

Figure Lengend Snippet: Comparison of the anticancer effects of first- (gefitinib) and second-generation EGFR-TKI (afatinib) in T790M mutation-negative oral cancer cell lines. A . Western blot analysis of p-EGFR and EGFR protein expressions in oral cancer cell lines treated with either gefitinib (8 µM) or afatinib (8 µM) for 24 h. Actin was used as the loading control. B . Trypan blue exclusion assay for cell viability. C . Annexin V/PI double staining to quantify the proportion of apoptotic cells. D . Western blot analysis of c-PARP and Mcl-1 protein expressions. All graphs are expressed as the mean ± SD of three independent experiments. * p < 0.05 by one-way ANOVA

Article Snippet: The following primary antibodies were used in the experiments: rabbit antihuman polyclonal antibodies against cleaved PARP (Cat. No. 9541; 1:1000), cleaved caspase-3 (Cat. No. 9664; 1:1000), phospho-mTOR (Cat. No. 2971; 1:1000), phospho-p70S6 (Cat. No. 9205; 1:1000), Bim (Cat. No. 2819; 1:1000), Mcl-1 (Cat. No. 4572; 1:1000), and rabbit antihuman monoclonal antibodies against phospho-EGFR (Cat. No. 3777; 1:1000), mTOR (Cat. No. 2983; 1:1000), and c-MYC (Cat. No. 5605; 1:1000) that were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA); and rabbit antihuman polyclonal antibody against EGFR (Cat. No. sc-03; 1:1000), goat antihuman polyclonal antibody against t-Bid (Cat. No. 34,325; 1:1000), and mouse antihuman monoclonal antibodies against β-actin (Cat. No. 47,778; 1:3000) and GAPDH (Cat. No. ab9484; 1:3000) that were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA).

Techniques: Comparison, Mutagenesis, Western Blot, Control, Trypan Blue Exclusion Assay, Double Staining

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: Antitumor activity of afatinib in EGFR T790M-negative human oral cancer therapeutically targets mTOR/Mcl-1 signaling axis

doi: 10.1007/s13402-024-00962-6

Figure Lengend Snippet: In vivo antitumor property of afatinib using a xenograft mouse model bearing HSC-3 cell line. Afatinib (25 and 50 mg/kg/day) was orally administered five times per week for 21 days to nude mice implanted with the HSC-3 cell line. A . Images of tumor specimens. B . Measurement of tumor volume. C . IHC analysis of tumor lysates for p-EGFR, Mcl-1, and c-caspase 3 (magnification, ×400). Measurement of body ( D ) and organ (liver and kidney) weights ( E ). F . H&E staining of tissues from control- and afatinib-treated mice. All graphs are expressed as the mean ± SD. * P < 0.05 by nonparametric Mann–Whitney tests

Article Snippet: The following primary antibodies were used in the experiments: rabbit antihuman polyclonal antibodies against cleaved PARP (Cat. No. 9541; 1:1000), cleaved caspase-3 (Cat. No. 9664; 1:1000), phospho-mTOR (Cat. No. 2971; 1:1000), phospho-p70S6 (Cat. No. 9205; 1:1000), Bim (Cat. No. 2819; 1:1000), Mcl-1 (Cat. No. 4572; 1:1000), and rabbit antihuman monoclonal antibodies against phospho-EGFR (Cat. No. 3777; 1:1000), mTOR (Cat. No. 2983; 1:1000), and c-MYC (Cat. No. 5605; 1:1000) that were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA); and rabbit antihuman polyclonal antibody against EGFR (Cat. No. sc-03; 1:1000), goat antihuman polyclonal antibody against t-Bid (Cat. No. 34,325; 1:1000), and mouse antihuman monoclonal antibodies against β-actin (Cat. No. 47,778; 1:3000) and GAPDH (Cat. No. ab9484; 1:3000) that were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA).

Techniques: In Vivo, Staining, Control, MANN-WHITNEY

Journal: Biomedicines

Article Title: Cetuximab-Toxin Conjugate and NPe6 with Light Enhanced Cytotoxic Effects in Head and Neck Squamous Cell Carcinoma In Vitro.

doi: 10.3390/biomedicines12050973

Figure Lengend Snippet: Figure 1. The expression levels of the EGFR protein and mRNA in each HNSCC cell line. WB of EGFR and GAPDH. EGFR protein was detected in Sa3, HO-1-u-1, and SAS, whereas HSQ-89 expression was nondetectable (a). EGFR mRNA expression measured by real −time PCR. Higher ∆Ct indicates lower EGFR mRNA expression. mRNA expression of EGFR was moderate in HO-1-u-1 and SAS, and high in Sa3. HSQ-89 showed a very low expression of EGFR. The red x−mark indicates the mean, and error bars show 95% confidence interval of n = 5 experiments (b). In the DepMap 23Q4 public dataset (https://doi.org/10.25452/figshare.plus.24667905.v2, accessed on 1 March 2024), the effect of EGFR gene knockout is shown in CHRONOS score against EGFR mRNA expression in HNSCC cell lines. Sa3 is absent from the dataset (c).

Article Snippet: After blocking, the membranes were incubated with primary rabbit polyclonal antibody against EGFR (dilution 1/1000, HPA018530, Atlas Antibodies, Stockholm, Sweden) and GAPDH (dilution 1/5000, 10494-1-AP, Proteintech, Rosemont, IL, USA) overnight at 4 ◦C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Gene Knockout

Journal: Biomedicines

Article Title: Cetuximab-Toxin Conjugate and NPe6 with Light Enhanced Cytotoxic Effects in Head and Neck Squamous Cell Carcinoma In Vitro.

doi: 10.3390/biomedicines12050973

Figure Lengend Snippet: Figure 5. Cell viability of IT-EGFR + PS + illumination compared with anti EGFR antibody + PS + illumination treated in each cell line. We compared the cytotoxicity of Cmab with PDT and IT-Cmab with PDT (iTAP) in Sa3 and HO-1-u-1 cells. The cytotoxicity of Cmab with PDT has no effect on Sa3 and HO-1-u-1 (a,b). On the other hand, the cytotoxicity of IT-Cmab with PDT (iTAP) was the IT dose-dependent in Sa3 and HO-1-u-1 cells (a,b).

Article Snippet: After blocking, the membranes were incubated with primary rabbit polyclonal antibody against EGFR (dilution 1/1000, HPA018530, Atlas Antibodies, Stockholm, Sweden) and GAPDH (dilution 1/5000, 10494-1-AP, Proteintech, Rosemont, IL, USA) overnight at 4 ◦C.

Techniques:

Journal: Scientific Reports

Article Title: A single mutation in the E2 glycoprotein of hepatitis C virus broadens the claudin specificity for its infection

doi: 10.1038/s41598-022-23824-3

Figure Lengend Snippet: HCV-JFH1-tau Lot B1 having M706L mutation can infect non-hepatic iPS cells via the CLDN6-dependent pathway. ( a ) Cellular expression patterns of various HCV entry factors. Huh7.5.1-8 and 253G1 cells were lysed, and equal protein amounts of each cell lysate (10 μg) were subjected to immunoblotting for the SRBI, EGFR, LDLR, CLDN1, CLDN6, OCLN, CD81, and GAPDH proteins. ( b ) Huh7.5.1-8 (squares), OKH-4 (triangles), and 253G1 (diamonds) cells were infected with HCV-JFH1-tau or HCV-JFH1-tau Lot B1 at 1.0 × 10 5 Geq/cell. At 0.5, 1, 1.5, and 2 days p.i., cellular HCV RNA contents were measured by qRT-PCR. Values are expressed as the ratio of OKH-4 at each day p.i. Data are presented as the mean ± S.D. (n = 6). *, p < 0.01 (vs. values of OKH-4 cells at each time point; Student’s t test). ( c ) 253G1 cells and Huh7.5.1-8 cells were infected with HCV-JFH1-tau Lot B1 at 1.0 × 10 5 Geq/cell in the presence of DMSO (white) or 1 μM sofosbuvir, a direct-acting antiviral agent (DAA, black). At 2 days p.i., HCV RNA contents in cells and culture supernatants were measured by qRT-PCR. Data are presented as the mean ± S.D. (n = 10 in 253G1 and n = 6 in Huh7.5.1-8). *, p < 0.01 (vs. values of DMSO treated cells; Student’s t test). ( d ) 253G1 cells (white) and S7-A cells (black) were preincubated with 5.0 μg/ml of control mouse IgG, or 5.0 μg/ml of anti-CD81 mAb (clone JS-81), 5.0 μg/ml of anti-CLDN6 mAb (clone 342927), 3.0 μg/ml of control rat IgG, or 3.0 μg/ml of mAb against OCLN (clone 1–3) for 30 min at room temperature and then infected with HCV-JFH1-tau Lot B1 at 1.0 × 10 5 Geq/cell. At 4 days p.i., cellular HCV RNA contents were measured by qRT-PCR. Values are expressed as the percentage of control mouse or rat IgG. Data are presented as the mean ± S.D. (n = 6).

Article Snippet: Rabbit polyclonal antibodies against EGFR were purchased from Cell signaling (Massachusetts, USA).

Techniques: Mutagenesis, Expressing, Western Blot, Infection, Quantitative RT-PCR, Control