Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: SRC-3 +/+ ApoE -/- mice exhibit more severe atherosclerosis. (A) SRC-3 was highly expressed in human atherosclerotic plaques. N represents plaque-adjacent vasculature in the lower limb aorta. AS represents atherosclerotic plaques in the lower limb aorta. (B) SRC-3 expression was upregulated in the aortas of ApoE -/- mice after the mice were fed a WD for 12 weeks. (C) SRC-3 was expressed in the endothelial cells and vascular smooth muscle cells of chow-fed SRC-3 -/- ApoE -/- mice and was further increased after the mice were fed a WD for 12 weeks. Sections of frozen aortic roots from SRC-3 +/+ ApoE -/- and SRC-3 -/- ApoE -/- mice were subjected to X-gal staining. Arrows indicate positively stained cells (blue). Scale bar, 100 µm. (D) SRC-3 promoted atherosclerotic plaque formation. Representative images of en face Oil Red O-stained aortas from SRC-3 +/+ ApoE -/- and SRC-3 -/- ApoE -/- mice (left panel). Quantification of the plaque areas in aortas (right panel). (E-K) Cross-sections of the aortic roots from SRC-3 +/+ ApoE -/- and SRC-3 -/- ApoE -/- mice were subjected to (E-F) H&E staining (scale bar, 500 µm (E); scale bar, 200 µm (F)), (G) α-SMA staining (scale bar, 200 µm), (H-I) Masson staining (scale bar, 200 µm (H); scale bar, 200 µm (I)), (J) Oil Red O staining (scale bar, 500 µm), and (K) F4/80 staining (scale bar, 200 µm). Left panels, representative images. Right panels, quantification of stained area or a percentage of lesion area. “×” indicates necrotic area. (L) Plaque stability was significantly increased in SRC-3 -/- ApoE -/- mice. The data represent the mean ± SEM. The results are representative of three independent experiments. P values were calculated by unpaired two-tailed Student's t-test. *, P <0.05; ** P <0.01.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Expressing, Staining, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: SRC-3 in endothelial cells contributes to the development of atherosclerosis. (A) Western blot showing that AAV-mediated SRC-3 shRNA decreased SRC-3 expression levels in the aortas of ApoE -/- mice. (B) SRC-3 knockdown reduced WD-induced atherosclerotic plaque formation in ApoE -/- mice. Representative images of en face Oil Red O-stained aortas from ApoE -/- mice injected with AAV-mediated SRC-3 shRNA and scramble shRNA (left panel). Quantification of the plaque areas of aortas (right panel). The data represent the mean ± SEM. (C-F) Cross-sections of the aortic roots of ApoE -/- mice injected with AAV-mediated SRC-3 shRNA and scramble shRNA were subjected to (C) H&E staining (scale bar, 100 µm), (D) Masson staining (scale bar, 500 µm), (E) Oil Red O staining (scale bar, 100 µm), (F) F4/80 staining (scale bar, 100 µm). Left panels, representative images. Right panels, quantification of stained area or a percentage of lesion area. The data represent the mean ± SEM. The results are representative of three independent experiments. P values were calculated by unpaired two-tailed Student's t-test. *, P <0.05; **, P <0.01; ***, P <0.001.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Western Blot, shRNA, Expressing, Staining, Injection, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: SRC-3 increases ICAM-1 expression during atherosclerosis development. (A) KEGG enrichment pathway analysis and (B) Gene Ontology (GO) biological process analysis of mRNA profiles in the aortas of SRC-3 +/+ ApoE -/- and SRC-3 -/- ApoE -/- mice after WD feeding for 12 weeks. (C) Selected genes involved in leukocyte recruitment and proinflammatory markers are shown as a heat map. (D) The mRNA level of ICAM-1 in the aortas of SRC-3 -/- ApoE -/- mice was significantly decreased after WD feeding for 12 weeks. (E) The protein level of ICAM-1 in the aortas of SRC-3 +/+ ApoE -/- and SRC-3 -/- ApoE -/- mice after WD feeding for 12 weeks. Each lane represents a pooled sample of three representative mice. (F) Western blot analysis of SRC-3 and ICAM-1 in 16 atherosclerotic plaques and plaque-adjacent vasculature in the lower limb aorta of accident patients. N represents plaque-adjacent vasculature in the lower limb aorta; AS represents atherosclerotic plaques in the lower limb aorta. (G) Correlation between SRC-3 and ICAM-1 protein levels in 16 atherosclerotic plaques and plaque-adjacent vasculature in the lower limb aorta. The data represent the mean ± SEM. The results are representative of three independent experiments. P values were calculated by unpaired two-tailed Student's t-test. *, P <0.05.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Expressing, Western Blot, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: SRC-3 regulates ICAM-1 expression via enhancing NF-κB signaling. (A) The protein levels of SRC-3 and ICAM-1 in SRC-3 siRNA-transfected HUVECs were significantly reduced compared with those in scrambled siRNA-transfected HUVECs after TNFα or IL-1β treatment. (B) The mRNA level of ICAM-1 in the SRC-3 siRNA-transfected HUVECs was markedly decreased after TNFα or IL-1β treatment. (C) ICAM-1 promoter activity was reduced in SRC-3 siRNA-transfected HUVECs after TNFα or IL-1β treatment. (D) SRC-3 cooperated with p65 to enhance the activity of the NF-κB reporter (upper panel) and ICAM-1 promoter (lower panel). (E) NF-κB binding site mutation abolished NF-κB-mediated ICAM-1 promoter activity. (F) The recruitment of SRC-3 and p65 was significantly reduced in SRC-3 siRNA-transfected HUVECs after TNFα or IL-1β treatment (right panel). Position of the subfragments detected by ChIP assays (left panel). (G) The SRC-3 siRNA-transfected HUVECs monolayer exhibited a significantly decreased number of adhered (upper panel) and transmigrated (lower panel) THP-1 cells after TNFα or IL-1β treatment. (H) Western blot showing ICAM-1 overexpression in SRC-3 siRNA-transfected HUVECs. (I) ICAM-1 overexpression in SRC-3 siRNA-transfected HUVECs rescued monocyte attachment to HUVECs and monocyte transendothelial migration after TNFα or IL-1β treatment. The data represent the mean ± SEM of three independent experiments. P values were calculated by unpaired two-tailed Student's t-test. *, P <0.05; **, P <0.01;***, P <0.001.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Expressing, Transfection, Activity Assay, Binding Assay, Mutagenesis, Western Blot, Over Expression, Migration, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: Pharmacological inhibition of SRC-3 reduces atherosclerosis. (A) The protein levels of SRC-3, ICAM-1 and p-p65 in bufalin-treated HUVECs were significantly reduced compared with those in vehicle-treated HUVECs after TNFα or IL-1β treatment. The bufalin-treated HUVECs monolayer resulted in a dramatically decreased number of adhered (upper panel) and transmigrated (lower panel) THP-1 cells after TNFα or IL-1β treatment. (B-C) ApoE -/- mice were administered vehicle or bufalin (1.0 mg/kg, six times a week) by intraperitoneal injection for 13 weeks concomitant with WD feeding. (B) Dosing regimen (ApoE -/- prevention model). (C) Representative images of en face Oil Red O-stained aortas from ApoE -/- mice treated with vehicle or bufalin (left panel). Quantification of the plaque areas of aortas (right panel). The data represent the mean ± SEM. (D) The protein levels of SRC-3 and ICAM-1 in the aortas of ApoE -/- mice treated with bufalin were significantly reduced in the ApoE -/- prevention model. (E-F) ApoE -/- mice were fed a WD for 10 weeks and then treated with vehicle or bufalin (1.0 mg/kg, six times a week) by intraperitoneal injection for 13 weeks concomitant with WD feeding. (E) Dosing regimen (ApoE -/- regression model). (F) Representative images of en face Oil Red O-stained aortas from ApoE -/- mice treated with vehicle or bufalin (left panel). Quantification of the plaque areas of aortas (right panel). The data represent the mean ± SEM. (G) The protein levels of SRC-3 and ICAM-1 in the aortas of ApoE -/- mice treated with bufalin were significantly reduced in the ApoE -/- regression model. The data represent the mean ± SEM. The results are representative of three independent experiments. P values were calculated by unpaired two-tailed Student's t-test. *, P <0.05; **, P <0.01;***, P <0.001.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Inhibition, Injection, Staining, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: SRC-3 deficiency prevents atherosclerosis development by decreasing endothelial ICAM-1 expression to attenuate macrophage recruitment

doi: 10.7150/ijbs.74864

Figure Lengend Snippet: Schematic model of the mechanism by which SRC-3 accelerates atherosclerosis development. SRC-3 promotes atherosclerosis development by increasing ICAM-1 transcription by enhancing the function of NF-κB in endothelial cells to promote macrophage recruitment. SRC-3 depletion or pharmacological inhibition of SRC-3 by bufalin ameliorates atherosclerosis development through decreasing endothelial ICAM-1 expression and macrophage recruitment via reduction of NF-κB function.

Article Snippet: Anti-SRC-3 antibodies (C-20, sc-7216) and anti-p65 (D14E12, #8284) antibodies were purchased from Santa Cruz Biotechnology and Cell Signaling Technology, respectively.

Techniques: Inhibition, Expressing

Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Histology and immunohistochemistry (IHC) of ALCL tumor samples. (A, B) Histology of ALCL tumor samples (HE staining). (A) The classic type of ALCL. The tumor cells are large with abundant cytoplasm and manifest prominent nucleoli with an eccentrically located and pleomorphic nucleus of kidney-shape. (B) ALCL of small cell type. The tumor cells are small in size, with abundant cytoplasm and prominent nucleoli. (C to H) IHC staining of ALCL tumor samples (EnVision staining). Expression of ALK, p-AKT, p-mTOR, p-4E-BP1, and p-p70S6K eIF4E was assessed. (C, D) IHC staining with ALK1 antibody. (C) The tumor cells show cytoplasmic and nuclear staining. (D) The tumor cells manifest cytoplasmic staining only. (E) IHC staining with Thr308p-AKT antibody. The tumor cells show cytoplasmic and nuclear staining. (F) IHC staining with Ser2448p-mTOR antibody. The tumor cells show cytoplasmic staining. (G) IHC staining with Thr70p-4E-BP1 antibody. The tumor cells show cytoplasmic staining. (H) IHC staining with Thr421p-p70S6K antibody. The tumor cells show cytoplasmic staining. All images were captured at 400× magnification.

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Relationship between expression of p-4E-BP1 and p-p70S6K and activation of ALK, AKT, and mTOR

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Expressing, Activation Assay, Significance Assay

Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Overexpression of NPM-ALK activated the AKT/mTOR pathway. Overexpression of NPM-ALK in BaF3 cells induced hyper-activation of the AKT/mTOR pathway, as demonstrated by hyperphosphorylation of AKT and downstream molecules of mTOR: 4E-BP1 and p70SK6.

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Over Expression, Activation Assay

Journal: Nutrition and cancer

Article Title: The effect of gartanin, a naturally- occurring xanthone in mangosteen juice, on the mTOR pathway, autophagy, apoptosis and the growth of human urinary bladder cancer cell Lines

doi: 10.1080/01635581.2013.785011

Figure Lengend Snippet: Schematic presentation of mechanisms of gartanin’s action in induction of autophagy and apoptosis. (1) Gartanin activates AMPK α and inhibits AKT activation, which causes down-regulation of p70S6 and 4E-BP1 leading to autophagy; (2) Gartanin down-regulates Bcl-2 and up-regulates p53, PUMA and Bax, which results in activation of Caspase-3 and PARP cleavages to induce apoptosis.

Article Snippet: Antibodies for phospho-AMP-activated protein kinase α (AMPKα), AMPKα, phospho-4E-BP1, 4E-BP1, phospho-rpS6, phospho-ACC, ACC, Caspase 3, PARP, phospho-mTOR, mTOR, TSC1, TSC2, p21, p70S6, phospho-p70S6, and p62 were purchased from Cell Signaling Technologies (Beverly, MA).

Techniques: Activation Assay

Journal: Frontiers in Endocrinology

Article Title: PKC-mediated phosphorylation and activation of the MEK/ERK pathway as a mechanism of acquired trastuzumab resistance in HER2-positive breast cancer

doi: 10.3389/fendo.2022.1010092

Figure Lengend Snippet: Selected antibodies, fluorescence intensities and Log 2 FC in protein & phosphoprotein quantities in T15 and JIMT-1 relative to SKBR3.

Article Snippet: PI3K/mTOR , 4E-BP1 - phosphoThr70 , Cell Signaling , 9455 , Rb pAb , 754 , 119 , 164 , 2.66 , 0.46.

Techniques: Fluorescence