anti insulin like growth factor 2 receptor igf2r  (Cell Signaling Technology Inc)

Journal: Cancer Research

Article Title: Integrative Multiomic Classification Reveals Distinct Origins and Evolutionary Trajectories of Thymic Epithelial Tumors

doi: 10.1158/0008-5472.CAN-24-4977

Figure Lengend Snippet: CN-type thymoma–specific IRS4 activation. A, Volcano plot showing differentially expressed genes between GTF2I-type and CN-type thymomas. The P values were calculated by a Student t test and adjusted with the Benjamini–Hochberg procedure. Known oncogenes (reported in COSMIC Cancer Gene Census) are highlighted in red. B, Box plot showing exclusive IRS4 expression in CN-type thymomas. The unit of the y -axis is TPM. The P values were calculated by the Wilcoxon test. C, Relationship between IRS4 expressions and IGF/PI3K/AKT cascade in CN-type tumors. IRS4 showed strong positive correlation with PI3K cascade score (REACT_976; top right), IGF receptor signaling pathway score (GO:0048009; top left), and their representative individual genes, such as IGFR1 (bottom left) and PIK3R1 (bottom right). Pearson correlation tests were conducted. D, Western blot results comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 in HEK293T cells results in an increase of AKT and phosphorylated AKT (pAKT), implying activation of the PI3K/AKT pathway. The activation of the PI3K/AKT pathway was sustained in a serum-free condition, which further supports the role of IRS4 in the pathway activation. E, Cell proliferation assay comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 increases cell proliferation rate of HEK293T cells. F, Cell viability assay comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 decreases response to OSI-906 (IGF1R inhibitor). G, Kaplan–Meier curves of recurrence-free survival showing worse prognosis of IRS4 -high CN-type thymomas (red) than GTF2I-type (blue) or IRS 4-low CN-type thymomas (orange). The median value of IRS4 expression (TPM = 30) separates CN-type thymomas into IRS4 -high or -low subgroups. The P values were calculated by the log-rank test. H, Percentage of TET cases harboring driver mutations ( HRAS , GTF2I , and CYLD ) or showing IRS4 activation in each histologic type.

Article Snippet: An insulin-like growth factor 1 receptor (IGF1R) inhibitor, linsitinib (OSI-906), was purchased from Selleckchem.

Techniques: Activation Assay, Expressing, Western Blot, Control, Over Expression, Proliferation Assay, Viability Assay

Journal: PLOS One

Article Title: Cellular repressor of E1A-stimulated genes 1 enhances skeletal muscle performance through the stimulation of muscle differentiation and Akt-mTOR signaling pathway activation

doi: 10.1371/journal.pone.0328485

Figure Lengend Snippet: (A) Western blotting for CREG1 was performed using the serum (1 µL) of wild type (WT) and Tg mice. (B-C) The expression of CREG1 in the soleus muscles of WT and Tg mice was examined using RT-PCR and western blot analysis. Relative levels of Creg1 mRNA (B) and CREG1 protein (C) in the soleus muscles. (D) Relative levels of insulin-like growth factor 2 receptor (IGF2R), phosphorylated Akt Ser 473 (pAkt), total Akt (Akt), phosphorylated mechanistic target of rapamycin (pmTOR) Ser 2448 and total mTOR (mTOR) in the soleus muscles. Data were normalized to those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). (E) Western blotting for IGF2R, pAkt, Akt, pmTOR, mTOR and GAPDH in the soleus muscles. Representative images are shown. Data are presented as mean ± SEM; n = 4–6 per group. A student's t -test was performed; * P < 0.05, versus WT.

Article Snippet: The membranes were blocked for 1 h at room temperature in nonfat dry milk and then incubated overnight at 4 °C with the following antibodies: anti-CREG1 (ab233282; Abcam, Cambridge, UK), anti-Akt Ser 473 [9271; Cell Signaling Technology (CST), Danvers, MA, USA], anti-Akt (9272; CST), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2118; CST), anti- insulin-like growth factor 2 receptor (IGF2R) (14364; CST), anti- mTOR Ser 2448 (2971; CST), anti- mTOR (2972; CST), and anti-α/β-tubulin (2148; CST).

Techniques: Western Blot, Expressing, Muscles, Reverse Transcription Polymerase Chain Reaction