Human HB‑EGF Antibody

Detection of Mouse HB-EGF by Western Blot

Dynamic expression and processing of Tgf-alpha and Hb-egf during multistep pancreatic neuroendocrine tumorigenesis (PNET). (A) Relative expression of Egf family members in cDNAs prepared from total RNA extracts of successive stages of neoplastic progression (NI = normal islets; HI = hyperplastic islets; AI = angiogenic islets; IT = islet tumors). Levels of mRNAs are expressed as a percentage of the mGus control mRNA. (B) Western blot analysis of the various forms of Tgf-alpha polypeptides in total protein extracts from successive stages of disease progression. (C) Analogous Western blot analysis of the various forms of Hb-egf polypeptides in total protein extracts from the successive disease stages. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/20975924), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human HB-EGF by Western Blot

BHLHE40-knockdown reduced hypoxia-induced expression of a panel of cytokines and growth factors. a Heatmaps of cytokines and growth factors whose hypoxia-induced expression (1% O2 at 6 h or 48 h, fold-change ≥ 1.5 in two independent experiments) was diminished by BHLHE40-knockdown (KD) in LM cells. The gene expression levels were determined using the Illumina Human HT-12 expression BeadChips. Normalized (quantile normalization) hybridization signals were log2 transformed and standardized by genes across experiment conditions to generate the heatmap. b Heatmaps of a subset of genes list in a whose expression was affected by BHLHE40-KD in LM cells exposed to hypoxia combined with low (1 mM) glucose (1%O2/LG, 4 h). The gene expression levels were determined using the Affymetrix Human Gene 1.0 ST array. c Heatmaps of hypoxia-induced genes whose expression was not significantly affected by BHLHE40-KD in LM cells as determined by the Illumina Human HT-12 expression BeadChips. d Expression of luciferase reporters driven by hypoxia-responsive elements of ITGA6 or LDHA was not affected by BHLHE40 knockout (KO) by CRISPR/Cas9 editing in MDA-MB-231 cells, in the absence or presence of exogenous HIF1A. Luciferase activities were normalized to co-transfected CMV-beta -galactosidase and presented as mean ± SD (n = 6). e Expression of genes in control LM empty vector (EV) and LM BHLHE40-KD cells exposed to 1%O2/LG (4 h). mRNA expression levels were determined by qPCR, normalized to RPL13A, and presented as mean ± SD (n = 6). *p <  0.05 (n = 6, 1%O2/LG vs. untreated control), **p <  0.05 (n = 6, KD vs. EV), one-way ANOVA followed by Tukey’s post-hoc tests. f mRNA and protein expression levels of HBEGF and CTGF in primary xenograft tumors, determined by qPCR and immunoblotting, respectively. *p <  0.05 (n = 6, KD vs. EV), Student’s t test. Representative immunoblotting images of three tumors of KD or EV cells are presented Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30285805), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse HB-EGF by Western Blot

Egfr activity contributes to RT2 tumor growth and angiogenic switching. (A) Relative expression quantified by real-time quantitative PCR of Erbb family members in cDNAs derived from total RNA extracts of successive stages of pancreatic neuroendocrine tumorigenesis from RIP1-Tag2 (RT2) transgenic mice (NI = normal islets; HI = hyperplastic islets; AI = angiogenic islets; IT = islet tumors). Levels of mRNAs are expressed as a percentage of the mGus control mRNA. (B) Western blot analysis of protein extracts from successive stages of RT2-derived lesions. (C) Western blot analysis of protein extracts from RT2-derived tumors 4 h after treatment with a vehicle solution or erlotinib (80 mg/kg). (D-F) Comparison of the average tumor burden of RT2 mice treated daily with a control solution (vehicle) or with (D) gefitinib (80 mg/kg), (E) CI-1033 (80 mg/kg), both from 11.5 to 14.5 weeks of age, or with (F) erlotinib (80 mg/kg), from 12 to 16 weeks of age with an additional vehicle-treated time point at 14 weeks. (G) Comparison of the average number of hemorrhagic angiogenic islets per pancreas of RT2 mice treated daily with a vehicle solution or with erlotinib (80 mg/kg) or CI-1033 (80 mg/kg) from 6 to 9 weeks of age. (N = number of animals per treatment group). *P < 0.01. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/20975924), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse HB-EGF by Western Blot

Hb-egf contributes to the angiogenic switch and neovascularization of PNET tumors. (A) Comparison of the average number of hemorrhagic angiogenic islets in wild-type (wt) and Hb-egf mutant (Hb) 9-week-old RT2 mice. (B-C) Comparison of the average (B) tumor burden or (C) tumor number in wild-type and Hb-egf mutant 14-week-old RT2 mice. (D) Western blot analysis of Hb-egf protein expression in total protein extracts from pools of 5 wild-type or 5 Hb-egf mutant tumors. (E-F) Average percentage of (E) dividing tumor cells (phospho-histone H3 positive) or (F) apoptotic cells (TdT-mediated dUTP-biotin nick end-labeling [TUNEL] positive) in wild-type or Hb-egf mutant tumors at 14 weeks of age. (G) Average vessel density (ratio of Meca-32 stained area to total section area) in wild-type or Hb-egf mutant tumors at 14 weeks of age. (H-I) Representative micrographs of (H) wild-type or (I) Hb-egf mutant RT2 tumors stained with Meca-32 (red) and DAPI (blue) (200x); panels are representative of 20 wild-type and 18 Hb-egf RT2 tumors dissected from at least 5 independent mice of each genotype. (N = number of independent mice [A-C] or tumor fields [E-G] analyzed per genotype; 1-2 fields per tumor.) *P < 0.001. **P < 0.01. ***P < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20975924), licensed under a CC-BY license. Not internally tested by R&D Systems.