Journal: Journal of Cellular and Molecular Medicine

Article Title: MFTZ-1 reduces constitutive and inducible HIF-1α accumulation and VEGF secretion independent of its topoisomerase II inhibition

doi: 10.1111/j.1582-4934.2009.00822.x

Figure Lengend Snippet: Effects of MFTZ-1 on Akt and MAPK pathways in MDA-MB-231 cells. Cells were exposed to MFTZ-1 for 6 hrs at hypoxia (A), pre-starved MDA-MB-231 cells were exposed to MFTZ-1 for 4 hrs plus pre-stimulation with EGF (50 ng/ml) for 4 hrs at normoxia (B) before Western blotting analyses were done for the protein levels of HIF-1α, p-Akt, p-Erk1/2, p-p70s6k and p-4EBP1, Akt and Erk1/2. Data shown were representative of three independent experiments. (C) This scheme shows the relationship of Akt, Erk, p70s6k and 4EBP.

Article Snippet: The primary antibodies for p-Erk (#9101), Erk (#9102), p-Akt (#9272), Akt (#9271), p-4EBP1 (#9455) and p-p70S6K (#9206) were purchased from Cell Signaling Technology (Danvers, MA, USA), respectively.

Techniques: Western Blot

Journal: Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer

Article Title: Measles vaccine strains for virotherapy of non-small cell lung carcinoma

doi: 10.1097/JTO.0000000000000214

Figure Lengend Snippet: A) H2009, A549, and H838 cell lysates were assayed by immunoblot before and 48 hours after infection with MV-GFP for key proteins involved in cap-dependent and independent translation initiation. B) Beas2B expressing empty vector (Beas2B-V) and Beas2B expressing eIF4E (Beas2B-E) were infected with MV-CEA at indicated MOI and assayed for cell viability 72 hours after infection. 5’ cap-affinity assay of untreated Beas2B-V and Beas2B-E are shown. Relative increases in eIF4G binding in the cap-affinity assay correspond to enhanced eIF4F cap-complex formation. C) H2009 and H522 NSCLC cells were treated with MV-CEA (MOI=0.01), rapamycin (Rap), or the combination and assayed for cell viability 72 hours after infection. Supernatants were collected from cells and assayed for CEA production as a measure of viral replication. Error bars indicate standard deviation of the mean. * indicates statistical significance. D) H2009 and H522 cells were treated with MV-CEA (MOI=0.01), 4EGI-1 (15μM), or the combination and cell viability assayed 72 hours after infection. E) NSCLC cell lines were treated with MV-GFP at MOI of 0.25 either alone or in combination with rapamycin. Representative fluorescence micrographs are shown.

Article Snippet: The primary antibodies employed were rabbit α-eIF4E, rabbit α-4E-BP1, rabbit α-PARP, α-eIF2α, α-phospho-eIF2α(Ser 51 ), α-PKR, α-phospho-PKR, all from Cell Signaling at a 1:1000 dilution, mouse α-b-actin (Sigma) at a 1:10,000 dilution and rabbit α-eIF4GI (kindly provided by Nahum Sonenberg, McGill University Montreal, Quebec, Canada) at a 1:2500 dilution.

Techniques: Western Blot, Infection, Expressing, Plasmid Preparation, Binding Assay, Standard Deviation, Fluorescence

Journal: Physiological Reports

Article Title: Impact of resistance exercise on ribosome biogenesis is acutely regulated by post‐exercise recovery strategies

doi: 10.14814/phy2.12670

Figure Lengend Snippet: Effect of resistance exercise on the p38‐ MNK 1‐ eIF 4E axis. Phosphorylation levels of p38 (A), MNK 1 (B), and eIF 4E (C), and total levels of eIF 4E (D), total cyclin D1 protein (E), and mRNA levels (F). Representative Western blot figures (closest molecular weight marker is shown in the left‐hand side) (G). Western blot data were normalized to GAPDH , with the exception of eIF 4E Ser209, which was normalized to its respective total protein. Cyclin D1 mRNA expression was normalized to the geometric mean of three reference genes. Values are mean ± SEM . *different from PRE within the same trial ( P < 0.05), # different between trials within the same time point ( P < 0.05). Main effects and interactions are presented in the text.

Article Snippet: The following antibodies were used in this study: total UBF (#13125), p‐UBF Ser484 (#21638) and Ser388 (#21637), cyclin D1 (#450), total eIF4E (#9976) (Santa Cruz Biotechnology, Santa Cruz, CA), total TIF‐IA (#42539), p‐TIF Ser649 (#138651), GAPDH (#36840)(Abcam, Cambridge, U.K.), total Akt (#2920), p‐Akt Thr308 (#4056), total PRAS40 (#2691), p‐PRAS40 Thr246 (#2691), p‐p38 MAPK Thr180/Tyr182 (#4511), p‐eIF4E Ser209 (#9741), p‐MNK1 Thr197/202 (#2111)(Cell Signaling Technology, Inc., Danvers, MA), and total c‐Myc – 9E10 clone (Developmental Studies Hybridoma Bank, Iowa City, IA).

Techniques: Western Blot, Molecular Weight, Marker, Expressing