Journal: International journal of molecular sciences

Article Title: Cerebroprotective Effect of 17β-Estradiol Replacement Therapy in Ovariectomy-Induced Post-Menopausal Rats Subjected to Ischemic Stroke: Role of MAPK/ERK1/2 Pathway and PI3K-Independent Akt Activation.

doi: 10.3390/ijms241814303

Figure Lengend Snippet: Figure 6. Estradiol increases the CTMP protein expression. Carboxy-terminal modulator protein (CTMP) protein expression in brains from placebo- and E2-treated ovariectomized female rats sub- jected to tMCAO. (A) Representative Western blots and (B) relative abundance of CTMP in whole-cell lysates of non-ischemic (NI) and ischemic (I) hemispheres. Two-way ANOVA: E2 vs. placebo, p = 0.08, F1,24 = 3.1). Post hoc Sidak’s multiple comparisons test: significantly different from non-ischemic placebo group (* p < 0.05). Data are mean ± SEM of individual data points (normalized to β-actin). E2, 17β-estradiol. tMCAO, transient middle cerebral artery occlusion.

Article Snippet: Aliquots of protein (40 μg) were dissolved in NuPAGE LDS sample buffer (Invitrogen, Carlsbad, CA, USA) under reducing conditions, loaded on 4–12% Bis-Tris gels (Invitrogen), subjected to SDS-PAGE and electrotransferred to 0.2 μm nitrocellulose membranes for immunolabeling using the following primary antibodies: (1) anti-cleaved caspase-3 (Asp175), rabbit polyclonal antibody that detects endogenous levels of the large fragment (17/19 KDa) of activated caspase-3 (#9661;1:500; Cell Signaling Technology, Inc., Beverly, MA, USA); (2) anti-phospho MAPK (pERK1/2) rabbit monoclonal antibody, which recognizes ERK1 and ERK2 that are phosphorylated on both a Thr202 and a Tyr204 residue (D13.14.4E; #4370;1:2000; Cell Signaling Technology); (3) anti-MAPK1/2 (ERK1/2) rabbit polyclonal antibody (#06-182; 1:5000; Millipore, Temecula, CA, USA); (4) anti-phospho BAD (pBAD) rabbit monoclonal antibody, which detects endogenous levels of BAD only when phosphorylated at Ser136 (D25H8; #4366;1:500; Cell Signaling Technology); (5) anti-Bad rabbit polyclonal antibody (#9292; 1:500; Cell Signaling Technology); (6) anti-phospho Akt (pAkt) mouse monoclonal antibody, which recognizes Akt only when phosphorylated at Ser473 (193H12; #4058; 1:000; Cell Signaling Technology); (7) anti-Akt (Akt, pan) rabbit monoclonal antibody (C67E7; #4691; 1:1000, Cell Signaling Technology); (8) anti-CTMP, rabbit polyclonal antibody, which detects endogenous levels of total CTMP protein (#4612; 1:500; Cell Signaling Technology); and (9) anti-β-actin mouse monoclonal antibody (Clone AC-15, #A5441; 1:10000; Sigma, Saint Louis, MI, USA).

Techniques: Expressing, Western Blot

Journal:

Article Title: The Epstein-Barr Virus Immediate-Early Protein BZLF1 Induces Expression of E2F-1 and Other Proteins Involved in Cell Cycle Progression in Primary Keratinocytes and Gastric Carcinoma Cells

doi: 10.1128/JVI.76.24.12543-12552.2002

Figure Lengend Snippet: Representative cellular genes activated by BZLF1 expressiona

Article Snippet: Although BZLF1 was previously shown to induce a G 0 /G 1 block in HeLa cells ( 11 , 12 ), the results of the microarray experiment in telomerase-immortalized human keratinocyte cells suggested that BZLF1 is unlikely to induce a G 0 /G 1 cell cycle block in this cell type and instead may be activating cell cycle progression. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Gene Fold activation E2F-1 16 DP-1 43 Cyclin A 15 Cyclin E 35 Cdc25A 18 Thymidine kinase 5 Thymidylate kinase 21 Open in a separate window a RNA from telomerase-immortalized human keratinocytes, mock infected or infected with AdLacZ or AdBZLF1, was analyzed by microarray analysis using the Affymetrix GeneChip HuGeneFL Array.

Techniques: Activation Assay

Journal:

Article Title: The Epstein-Barr Virus Immediate-Early Protein BZLF1 Induces Expression of E2F-1 and Other Proteins Involved in Cell Cycle Progression in Primary Keratinocytes and Gastric Carcinoma Cells

doi: 10.1128/JVI.76.24.12543-12552.2002

Figure Lengend Snippet: BZLF1 increases the expression of E2F-1-responsive genes in some cell types. (A) Telomerase-immortalized keratinocytes or normal human fibroblasts were mock infected or infected with the AdLacZ or AdBZLF1 vector. Immunoblot analysis was performed 2 days later to quantitate expression of cyclin E, SLBP, or Cdc25A, as indicated. (B) AGS and primary tonsil keratinocytes were infected with the AdLacZ or AdBZLF1 vector, and immunoblot analysis was performed 2 days later to quantitate expression of cyclin E and SLBP as indicated.

Article Snippet: Although BZLF1 was previously shown to induce a G 0 /G 1 block in HeLa cells ( 11 , 12 ), the results of the microarray experiment in telomerase-immortalized human keratinocyte cells suggested that BZLF1 is unlikely to induce a G 0 /G 1 cell cycle block in this cell type and instead may be activating cell cycle progression. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Gene Fold activation E2F-1 16 DP-1 43 Cyclin A 15 Cyclin E 35 Cdc25A 18 Thymidine kinase 5 Thymidylate kinase 21 Open in a separate window a RNA from telomerase-immortalized human keratinocytes, mock infected or infected with AdLacZ or AdBZLF1, was analyzed by microarray analysis using the Affymetrix GeneChip HuGeneFL Array.

Techniques: Expressing, Infection, Plasmid Preparation, Western Blot