Journal: ACS Omega

Article Title: Topical Application of Baicalin Combined with Echinacoside Ameliorates Psoriatic Skin Lesions by Suppressing the Inflammation-Related TNF Signaling Pathway and the Angiogenesis-Related VEGF Signaling Pathway

doi: 10.1021/acsomega.3c04281

Figure Lengend Snippet: Transcriptome analysis of skin lesion tissues from Con and IMQ groups. (A) Representative pictures of dorsal skin and histopathological observation of skin lesions by HE staining (scale bar, 100 μm) between Con and IMQ groups after topical application of IMQ for 6 days. Two opposite arrows indicated the epidermis, and the red circle indicated the inflammatory cell infiltration. (B) KEGG pathway enrichment analysis of DEGs between Con and IMQ groups. (C) Gene set enrichment analysis (GSEA) of the TNF signaling pathway and the VEGF signaling pathway.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor (VRI) was purchased from MCE (New Jersey).

Techniques: Staining

Journal: ACS Omega

Article Title: Topical Application of Baicalin Combined with Echinacoside Ameliorates Psoriatic Skin Lesions by Suppressing the Inflammation-Related TNF Signaling Pathway and the Angiogenesis-Related VEGF Signaling Pathway

doi: 10.1021/acsomega.3c04281

Figure Lengend Snippet: Topical application of B2-E1 (BAI/ECH = 2:1) reduced the release of inflammatory factors including TNF-α, IL-17A, and IL-23, increased the anti-inflammatory cytokine IL-10, inhibited the angiogenesis-related cytokine VEGF-A and MMP-9, and improved the antioxidant ability better than the single component in IMQ-induced psoriatic skin lesions. (A) The levels of TNF-α, IL-17A, IL-23, and IL-10 in skin lesion tissues in each group ( n = 6). (B) The content of VEGF-A and MMP-9, the activity of TrxR, and the content of MDA in skin lesion tissues in each group ( n = 6). Data are represented as mean ± SD △△△ P < 0.001 versus the Con group, # P < 0.05, ## P < 0.01, and ### P < 0.001 versus the IMQ group, * P < 0.05, ** P < 0.01, and *** P < 0.001 versus the KB group, and ▲▲ P < 0.01 versus the B2-E1 group.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor (VRI) was purchased from MCE (New Jersey).

Techniques: Activity Assay

Journal: ACS Omega

Article Title: Topical Application of Baicalin Combined with Echinacoside Ameliorates Psoriatic Skin Lesions by Suppressing the Inflammation-Related TNF Signaling Pathway and the Angiogenesis-Related VEGF Signaling Pathway

doi: 10.1021/acsomega.3c04281

Figure Lengend Snippet: B2-E1 (BAI/ECH = 2:1) could notably regulate the mRNA expression of key genes in the TNF signaling pathway and the VEGF signaling pathway better than the single component in IMQ-induced mice. (A) B2-E1 could potently inhibit the mRNA expression of these genes related to key targets involved in the TNF signaling pathway better than the single component in IMQ-induced skin lesions ( n = 6). (B) B2-E1 could significantly decrease the mRNA expression of these genes related to key targets involved in the VEGF signaling pathway better than the single component in IMQ-induced skin lesions ( n = 6). Data are expressed as mean ± SD △ P < 0.05, △△ P < 0.01, and △△△ P < 0.001 versus the Con group, # P < 0.05 and ## P < 0.01 and ### P < 0.001 versus the IMQ group, * P < 0.05, ** P < 0.01, and *** P < 0.001 versus the KB group, and ▲ P < 0.05 and ▲▲ P < 0.01 versus the B2-E1 group.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor (VRI) was purchased from MCE (New Jersey).

Techniques: Expressing

Journal: ACS Omega

Article Title: Topical Application of Baicalin Combined with Echinacoside Ameliorates Psoriatic Skin Lesions by Suppressing the Inflammation-Related TNF Signaling Pathway and the Angiogenesis-Related VEGF Signaling Pathway

doi: 10.1021/acsomega.3c04281

Figure Lengend Snippet: Potential mechanism of topical application of BAI combined with ECH ameliorating psoriatic skin lesions by inhibiting the TNF signaling pathway and the VEGF signaling pathway. BAI may inhibit the expression of key cytokines and genes such as TNF-α, IL-17A, IL-23, and AKT1 by regulating the TNF signaling pathway, and ECH could inhibit the expression of key cytokines and genes such as VEGFA, MMP9, SRC, and NOS3 by regulating the VEGF signaling pathway.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor (VRI) was purchased from MCE (New Jersey).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: ALW-II-41-27 intestinal dysmotility in Trichinella spiralis -infected mice. Effects of ALW-II-41-27 on body weight (A) , fecal water content (B) , gastrointestinal motility (C) and AWR scores (D) in Trichinella spiralis -infected mice. The results are expressed as the mean ± SD ( n = 10 per group); # indicates a significant difference from control group ( P < 0.05); ## indicates a significant difference from control group ( P < 0.01); ∗ indicates a significant difference from PI-IBS group ( P < 0.05); ∗∗ indicates a significant difference from PI-IBS group ( P < 0.01).

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Infection, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: ALW-II-41-27 suppressed oxidative stress in Trichinella spiralis -infected mice. Effects of ALW-II-41-27 on the levels of serum 4-HNE (A) , serum protein carbonyl (B) , serum 8-OHdG (C) , serum HO-1 activity (D) , colonic 4-HNE (E) , colonic protein carbonyl (F) , colonic 8-OHdG (G) , colonic HO-1 activity (H) . The results are expressed as the mean ± SD ( n = 10 per group); ## indicates a significant difference from control group ( P < 0.01); ∗ indicates a significant difference from PI-IBS group ( P < 0.05); ∗∗ indicates a significant difference from PI-IBS group ( P < 0.01).

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Infection, Activity Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: ALW-II-41-27 inhibited inflammation in Trichinella spiralis -infected mice. Effects of ALW-II-41-27 on the expression of serum TNF-α (A) , serum IL-6 (B) , serum IL-17 (C) , serum ICAM-1 (D) , serum IL-10 (E) and serum NF-κB activity (F) , colonic TNF-α (G) , colonic IL-6 (H) , colonic IL-17 (I) , colonic ICAM-1 (J) , colonic IL-10 (K) and colonic NF-κB activity (L) . The results are expressed as the mean ± SD ( n = 10 per group); ## indicates a significant difference from control group ( P < 0.01); ∗ indicates a significant difference from PI-IBS group ( P < 0.05); ∗∗ indicates a significant difference from PI-IBS group ( P < 0.01).

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Infection, Expressing, Activity Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: ALW-II-41-27 inhibited LPS-induced oxidative stress and inflammation in NCM460 cells. Effects of ALW-II-41-27 on cell viability (A) , LDH leakage (B) , cellular 4-HNE (C) , cellular protein carbonyl (D) , cellular 8-OHdG (E) , cellular HO-1 activity (F) , cellular TNF-α (G) , cellular IL-6 (H) , cellular IL-17 (I) , cellular ICAM-1 (J) , cellular IL-10 (K) and cellular NF-κB activity (L) . ## Indicates a significant difference from control cells ( P < 0.01); ∗ indicates a significant difference from LPS group ( P < 0.05); ∗∗ indicates a significant difference from LPS group ( P < 0.01).

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Activity Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: ALW-II-41-27 activated Nrf2 signaling whereas inhibited NF-κB signaling in Trichinella spiralis -infected mice. The protein expressions of EphA2, nuclear Nrf2, cytosol Nrf2, and nuclear NF-κB/p65, cytosol NF-κB/p65, Lamin B, and β-actin in colon tissue lysates were measured by western blotting and densitometry analysis. The results are expressed as the mean ± SD ( n = 10 per group); ## indicates a significant difference from control group ( P < 0.01); ∗∗ indicates a significant difference from PI-IBS group ( P < 0.01).

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Infection, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: EphA2 knock-out partially inhibited the effects of ALW-II-41-27 on LPS-treated cells. NCM460 cells were transfected to obtain clones knock-out for EphA2. Control cells and EphA2 KO cells were treated for 24 h with ALW-II-41-27 (100 ng/mL) and then treated with LPS (50 ng/mL) for 24 h. The protein expressions of EphA2, nuclear Nrf2, cytosol Nrf2, and nuclear NF-κB/p65, cytosol NF-κB/p65, Lamin B and β-actin in colon tissue lysates were measured by western blotting and densitometry analysis. The results are expressed as the mean ± SD ( n = 10 per group); ## indicates a significant difference from control cells ( P < 0.01); ∗∗ indicates a significant difference from LPS group ( P < 0.01); ΔΔ indicates a significant difference from control cells treated LPS; && indicates a significant difference from control cells treated with ALW-II-41-27 and LPS.

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Knock-Out, Transfection, Clone Assay, Control, Western Blot

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: EphA2 overexpression abolished the effects of ALW-II-41-27 on LPS-treated cells. NCM460 cells were transfected to obtain clones overexpressing for EphA2. Control cells and EphA2 overexpressing cells were treated for 24 h with ALW-II-41-27 and then treated with LPS (50 ng/mL) for 24 h. The protein expressions of EphA2, nuclear Nrf2, cytosol Nrf2, and nuclear NF-κB/p65, cytosol NF-κB/p65, Lamin B and β-actin in colon tissue lysates were measured by western blotting and densitometry analysis. The results are expressed as the mean ± SD ( n = 10 per group); ## indicates a significant difference from control cells ( P < 0.01); ∗∗ indicates a significant difference from LPS group ( P < 0.01); ΔΔ indicates a significant difference from control cells treated LPS; && indicates a significant difference from control cells treated with ALW-II-41-27 and LPS.

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Over Expression, Transfection, Clone Assay, Control, Western Blot

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: EphA2 overexpression abolished the protective effects of ALW-II-41-27 on LPS-induced oxidative stress and inflammation. Effects of EphA2 overexpression on cell viability (A) , LDH leakage (B) , cellular 4-HNE (C) , cellular protein carbonyl (D) , cellular 8-OHdG (E) , cellular HO-1 activity (F) , cellular TNF-α (G) , cellular IL-6 (H) , cellular IL-17 (I) , cellular ICAM-1 (J) , cellular IL-10 (K) and cellular NF-κB activity (L) . ## Indicates a significant difference from control cells ( P < 0.01); ∗ indicates a significant difference from LPS group ( P < 0.05); ∗∗ indicates a significant difference from LPS group ( P < 0.01); ΔΔ indicates a significant difference from control cells treated LPS; && indicates a significant difference from control cells treated with ALW-II-41-27 and LPS.

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Over Expression, Activity Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-κB Signaling Pathways

doi: 10.3389/fphar.2018.00272

Figure Lengend Snippet: EphA2 knock-out partially inhibited the protective effects of ALW-II-41-27 on LPS-induced oxidative stress and inflammation. Effects of EphA2 knock-out on cell viability (A) , LDH leakage (B) , cellular 4-HNE (C) , cellular protein carbonyl (D) , cellular 8-OHdG (E) , cellular HO-1 activity (F) , cellular TNF-α (G) , cellular IL-6 (H) , cellular IL-17 (I) , cellular ICAM-1 (J) , cellular IL-10 (K) and cellular NF-κB activity (L) . ## Indicates a significant difference from control cells ( P < 0.01); ∗ indicates a significant difference from LPS group ( P < 0.05); ∗∗ indicates a significant difference from LPS group ( P < 0.01); ΔΔ indicates a significant difference from control cells treated LPS; && indicates a significant difference from control cells treated with ALW-II-41-27 and LPS.

Article Snippet: ALW-II-41-27 (Purify ≥ 99.3%) were purchased from MedChem Express (Monmouth Junction, NJ, United States).

Techniques: Knock-Out, Activity Assay, Control

Journal: Biomimetics

Article Title: Developing New Peptides and Peptide–Drug Conjugates for Targeting the FGFR2 Receptor-Expressing Tumor Cells and 3D Spheroids

doi: 10.3390/biomimetics9090515

Figure Lengend Snippet: Determination of K D values from SPR Analysis.

Article Snippet: In our studies, SPR was used to investigate the binding interactions of the three peptides as well as that of the known FGF Receptor Tyrosine Kinase Inhibitor—CAS 192705-79-6 (Santacruz Biotechnologies, Dallas, TX, USA) with the kinase domain of FGFR2.

Techniques: Control

Journal: Experimental Physiology

Article Title: Platelet‐rich plasma affects gap junctional features in myofibroblasts in vitro via vascular endothelial growth factor (VEGF)‐A/VEGF receptor

doi: 10.1113/ep090052

Figure Lengend Snippet: FIGURE 1 Morphofunctional evaluation of NIH/3T3 fibroblast-to-myofibroblast transition. (a) Typical fibroblast (in PM, proliferation medium) impaled by the patch electrode (scale bar represents 20 μm). (b) Resting membrane potential (RMP, in millivolts; n = 4–8 cells; one-way ANOVA with Bonferroni’s correction, P = 0.074, d.f. = 56). Exact P-values of any comparison are reported in Table A1 for clarity. (c–m) Representative immunofluorescence analysis of α-smooth muscle actin (α-sma) and vinculin expression. Nuclei are counterstained with propidium iodide. Scale bars represent 10 μm. (n,o) Densitometric analysis of the fluorescence intensity (FI, in arbitrary units, a.u) of indicated markers. (p) Morphometric analysis of mean cell surface area. Abbreviations: Anti VEGF-A (vascular endothelial growth factor-A) Ab, anti-VEGF-A neutralizing antibodies; DM, differentiation medium; IgG, irrelevant isotype-matched IgG; KRN633, selective pharmacological VEGFR inhibitor; PM, proliferation medium; PRP, platelet-rich plasma; VEGF-A 2, 20, soluble VEGF-A 2 or 20 ng/mL. Values are the mean ± SD. Significance of differences in (n) (α-sma): *** versus PM (P < 0.0001); * versus PM (P = 0.05); §§§ versus DM (P < 0.0001); §§ versus DM (P = 0.007); ### versus DM+PRP (P < 0.0001); @@@ versus DM+DMSO (P < 0.0001); ˆˆˆ versus DM+PRP+DMSO (P < 0.0001); & versus DM+PRP+KRN (P = 0.019); ◦◦◦versus DM+PRP+IgG (P < 0.0001); + versus VEGF-A 2 (P = 0.033). Significance of differences in (o) (vinculin): *** versus PM (P < 0.0001); §§§ versus DM (P < 0.0001); § versus DM (P = 0.049); ### versus DM+PRP (P < 0.0001); @@@ versus DM+DMSO (P < 0.0001); ˆˆˆ versus DM+PRP+DMSO (P < 0.0001); ◦◦◦versus DM+PRP+IgG (P < 0.0001). In (n) and (o), one-way ANOVA with Tukey’s post hoc test, n = 60 regions of interest. Significance of differences in (p) (cell surface area, one-way ANOVA with Tukey’s post hoc test, n = 36 cells): *** versus PM (P < 0.0001); * versus PM (P = 0.026); §§§ versus DM (P < 0.0001); ### versus DM+PRP (P < 0.0001); @@@ versus DM+DMSO (P < 0.0001); ˆˆˆ versus DM+PRP+DMSO (P < 0.0001); ◦◦◦versus DM+PRP+IgG (P < 0.0001)

Article Snippet: In parallel, cells were cultured: (1) in DMplus a specific ATP-competitive inhibitor of VEGFR tyrosine kinase activity, KRN633 (SantaCruz Biotechnology, Santa Cruz, CA, USA; IC50 = 170 nM, Cat# sc-204379, Lot# B1810, dilution in DMSO, Sigma, Cat# 472301) in the absence or presence of PRP; (2) in DM in the presence of PRP plus mouse monoclonal anti-VEGF-A neutralizing antibodies (10 μg/mL; Sigma, Cat# V4758, Lot# 066K1428, RRID:AB_477621).

Techniques: Membrane, Comparison, Immunofluorescence, Expressing, Fluorescence, Clinical Proteomics

Journal: Experimental Physiology

Article Title: Platelet‐rich plasma affects gap junctional features in myofibroblasts in vitro via vascular endothelial growth factor (VEGF)‐A/VEGF receptor

doi: 10.1113/ep090052

Figure Lengend Snippet: FIGURE 2 Morphofunctional evaluation of human skin primary fibroblasts. (a) Typical human primary skin fibroblasts (in PM) impaled by a patch electrode (scale bar represents 10 μm). (b) Resting membrane potential (RMP, in millivolts; n = 4–7 cells; one-way ANOVA with Bonferroni’s correction, P = 0.007, d.f. = 29; **P < 0.01 and ***P < 0.001 vs. PM; §§P < 0.01 and §§§P < 0.001 vs. DM; ##P < 0.01 and ###P < 0.001 vs. DM+PRP) in the indicated experimental conditions. Exact P-values of any comparison are reported in Table A6 for clarity. (c,e,g,i,k,m) Representative immunofluorescence analysis of α-sma and Cx43 expression in human cells cultured as specified in Figure 1. Merge: superimposed DIC and fluorescence images (scale bar represents 25 μm). (d,f,h,j,l,n) Representative time course of Ij (in picoamperes) from cell pairs in the indicated experimental conditions. Abbreviations: α-sma, α-smooth muscle actin; Cx43, connexin 43; DIC, differential interference contrast; DM, differentiation medium; Ij, transjunctional current; KRN633, selective pharmacological VEGFR inhibitor; PM, proliferation medium; PRP, platelet-rich plasma; VEGF, vascular endothelial growth factor-A 2 ng/ml

Article Snippet: In parallel, cells were cultured: (1) in DMplus a specific ATP-competitive inhibitor of VEGFR tyrosine kinase activity, KRN633 (SantaCruz Biotechnology, Santa Cruz, CA, USA; IC50 = 170 nM, Cat# sc-204379, Lot# B1810, dilution in DMSO, Sigma, Cat# 472301) in the absence or presence of PRP; (2) in DM in the presence of PRP plus mouse monoclonal anti-VEGF-A neutralizing antibodies (10 μg/mL; Sigma, Cat# V4758, Lot# 066K1428, RRID:AB_477621).

Techniques: Membrane, Comparison, Immunofluorescence, Expressing, Cell Culture, Fluorescence, Clinical Proteomics

Journal: Breast Cancer Research : BCR

Article Title: erbB3 recruitment of insulin receptor substrate 1 modulates insulin-like growth factor receptor signalling in oestrogen receptor-positive breast cancer cell lines

doi: 10.1186/bcr3018

Figure Lengend Snippet: Western blot analysis . (a) Total and phosphorylated insulin-like growth factor type I receptor (IGF-IR), insulin receptor substrate 1 (IRS-1), erbB3, Akt and extracellular-signal regulated kinase 1/2 (ERK1/2) protein expression following incubation of MCF-7 and T47D cells in medium containing either the specific IGF-IR/IR tyrosine kinase inhibitor 4-anilino-5-bromo-2-[4-(2-hydroxy-3-( N , N -dimethylamino)propoxy)anilino]pyrimidine (ABDP) (1 μM) or appropriate vehicle control for 24 hours and subsequently challenged with increasing concentrations of heregulin β1 (HRGβ1) (0.1 to 10 ng/ml) or vehicle control for 5 minutes. Densitometric analysis of phosphorylated Akt and ERK1/2 protein levels in (b) MCF-7 and (c) T47D cells treated with HRGβ1 in the absence and presence of ABDP. The results are expressed as the means ± standard errors of the mean of at least three separate experiments from the 1 ng/ml HRGβ1-primed groups. (d) Western blot analysis of total IGF-IR, erbB3 and IRS-1 expression following immunoprecipitation (IP) with total IRS-1 antibody in MCF-7 and T47D cells incubated for 24 hours in medium containing ABDP (1 μM) or vehicle control. * P ≤ 0.05 versus HRGβ1 for both cell lines.

Article Snippet: To examine the effects of pharmacological blockade of IGF-IR, cells were incubated in phenol red-free (white) RPMI medium supplemented with 5% FCS and either the IGF-IR/IR tyrosine kinase inhibitor 4-anilino-5-bromo-2-[4-(2-hydroxy-3-( N , N -dimethylamino)propoxy)anilino]pyrimidine (ABDP) (1 μM in dimethyl sulphoxide, AstraZeneca, Macclesfield, UK) [ ] or appropriate vehicle control for 1 to 2 days.

Techniques: Western Blot, Expressing, Incubation, Control, Immunoprecipitation

Journal: Breast Cancer Research : BCR

Article Title: erbB3 recruitment of insulin receptor substrate 1 modulates insulin-like growth factor receptor signalling in oestrogen receptor-positive breast cancer cell lines

doi: 10.1186/bcr3018

Figure Lengend Snippet: Western blot analysis . (a) Total and phosphorylated insulin-like growth factor type I receptor (IGF-IR), insulin receptor substrate 1 (IRS-1), Akt, extracellular signal-regulated kinase 1/2 (ERK1/2) and β-actin protein expression following incubation of MCF-7 and T47D cells in medium containing either lipid and control siRNA (C si) mix (100 nM) or lipid and IRS si mix (100 nM) in the absence or presence of 4-anilino-5-bromo-2-[4-(2-hydroxy-3-( N , N -dimethylamino)propoxy)anilino]pyrimidine (ABDP) (1 μM) for 2 days and subsequently challenged with heregulin β1 (HRGβ1) (10 ng/ml) for 5 minutes. Data are representative of at least three separate experiments. The effects of 4-day treatment with HRGβ1 (10 ng/ml), ABDP (0.1 μM) or a combination of HRGβ1 and ABDP on the growth of (b) MCF-7 cells or (c) T47D cells. The results are expressed as the means ± standard errors of the mean of triplicate wells and are representative of at least three separate experiments. * P ≤ 0.01 versus control, † P ≤ 0.001 versus ABDP.

Article Snippet: To examine the effects of pharmacological blockade of IGF-IR, cells were incubated in phenol red-free (white) RPMI medium supplemented with 5% FCS and either the IGF-IR/IR tyrosine kinase inhibitor 4-anilino-5-bromo-2-[4-(2-hydroxy-3-( N , N -dimethylamino)propoxy)anilino]pyrimidine (ABDP) (1 μM in dimethyl sulphoxide, AstraZeneca, Macclesfield, UK) [ ] or appropriate vehicle control for 1 to 2 days.

Techniques: Western Blot, Expressing, Incubation, Control