Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: NTRK1-mediated protection against manganese-induced neurotoxicity and cell apoptosis via IGF2 in SH-SY5Y cells.

doi: 10.1016/j.biopha.2023.115889

Figure Lengend Snippet: Fig. 9. Si-IGF2 Downregulates UPR Expression in Various Conditions Under MnCl2 Treatment. (A-E) Real-time PCR Analysis: Quantification of ER stress-related genes (GRP78, PREK, IRE-1, ATF-6, XBP1-s) mRNA levels normalized to GAPDH in different conditions: control + MnCl2, NTRK1-OE + MnCl2, Si-IGF2 + MnCl2, and Si-IGF2- NTRK1-OE + MnCl2. (F) Western Blot Analysis: Representative Western blot illustrating the expression of p-PERK, p-IRE1, and GRP78 in the mentioned conditions. Protein bands corresponding to these markers are visible in each condition. (G) Quantification of GRP78/GAPDH Expression. (H) Quanti fication of p-PERK/PERK Expression. (I) Quantification of p-IRE1/IRE1 Expressions. Statistical significance indicated by asterisks (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001) using one-way ANOVA with Bonferroni post-test. Error bars indicate means ± SEM, n = 3.

Article Snippet: Membranes were incubated with 5% milk in Tris-buffered saline with Tween 20 (TBST, 0.1%) at routine temperature for 2 h. Then, incubated overnight at 4 ◦C with the following primary antibodies: rabbit anti–TRKA (1:1000; A4147; ABclonal, Wuhan, China), rabbit anti-GRP78 (1:2000; 11587–1-AP; Proteintech, Wuhan, China), rabbit anti-p-PERK (1:400; DF7576; Affinity, Jiangsu, China), rabbit anti- PERK (1:1000; 20582–1-AP; Proteintech, Wuhan, China), rabbit anti-p-IRE (1:400; AF7150; Affinity, Jiangsu, China), rabbit anti- IRE (1:1000; 27528–1-AP; Proteintech, Wuhan, China), rabbit anti-IGF2 (1:1000, ab9574, Abcam, USA), rabbit anti- cleaved caspase-3 (1:1000, 9664; Cell Signaling Technology, USA), and rabbit anti-GAPDH (1:5000; 10494–1-AP; Proteintech, Wuhan, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: Oncology Reports

Article Title: Synergistic activity of agents targeting growth factor receptors, CDKs and downstream signaling molecules in a panel of pancreatic cancer cell lines and the identification of antagonistic combinations: Implications for future clinical trials in pancreatic cancer

doi: 10.3892/or.2020.7822

Figure Lengend Snippet: The membrane bound expression level of various growth factor receptors determined by flow cytometry in human pancreatic cancer cell lines represented as histograms. EGFR, epidermal growth factor receptor; HER, human epidermal growth factor receptor; c-MET, mesenchymal-epithelial transition factor; IGF-IR, insulin-like growth factor 1 receptor; ALK7, anaplastic lymphoma kinase 7.

Article Snippet: The antibodies for flow cytometry including mouse anti-EGFR (HM43.16B) and anti-HER2 (HM50.67A) were raised in-house against the external domain of these receptors ( ) whereas mouse anti-HER3 (MAB3481), anti-HER4 (MAB11311), ALK7 (MAB77491), HGF R/c-MET (MAB3582), PDGFRα (MAB1264), PDGFRβ (MAB1263) and IGF-IR (MAB391) were purchased from R&D Systems (Europe Ltd. UK) and Insight Biotechnology (Middlesex, UK), respectively.

Techniques: Membrane, Expressing, Flow Cytometry

Journal: Oncology Reports

Article Title: Synergistic activity of agents targeting growth factor receptors, CDKs and downstream signaling molecules in a panel of pancreatic cancer cell lines and the identification of antagonistic combinations: Implications for future clinical trials in pancreatic cancer

doi: 10.3892/or.2020.7822

Figure Lengend Snippet: Effect of afatinib, dinaciclib, dasatinib, stattic and NVP-AEW742 with or without ligands (EGF, HB-EGF, IGF-II) on the phosphorylation of EGFR and downstream cell signaling molecules including MAPK, AKT, STAT3, SRC and IGF-IR in BxPC-3 (A) and Capan-1 (B) cells. The cells were cultured in 10% FBS RPMI-1640 medium to near confluency. Cells were washed once with 0.5% FBS RPMI-1640 medium and incubated with selected agents (400 nM) for 1 h and then stimulated with 40 nM ligands (EGF, HB-EGF and IGF-II) for 15 min. Cells were then lysed, separated using SDS-PAGE, transferred onto PDVF membranes, probed with the antibodies of interest and visualized using LI-COR software. EGF, epidermal growth factor; HB-EGF, heparin-binding EGF-like growth factor; IGF-II, insulin-like growth factor II; EGFR, epidermal growth factor receptor; MAPK, mitogen-activated protein kinase; AKT, protein kinase B or PKB; STAT3, signal transducer and activator of transcription 3; SRC, proto-oncogene tyrosine kinase SRC; IGF-IR, insulin-like growth fact) or 1 receptor.

Article Snippet: The antibodies for flow cytometry including mouse anti-EGFR (HM43.16B) and anti-HER2 (HM50.67A) were raised in-house against the external domain of these receptors ( ) whereas mouse anti-HER3 (MAB3481), anti-HER4 (MAB11311), ALK7 (MAB77491), HGF R/c-MET (MAB3582), PDGFRα (MAB1264), PDGFRβ (MAB1263) and IGF-IR (MAB391) were purchased from R&D Systems (Europe Ltd. UK) and Insight Biotechnology (Middlesex, UK), respectively.

Techniques: Phospho-proteomics, Cell Culture, Incubation, SDS Page, Software, Binding Assay