Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Acute Regulation of Translation Initiation by Gonadotropin-Releasing Hormone in the Gonadotrope Cell Line LβT2

doi: 10.1210/me.2003-0478

Figure Lengend Snippet: LβT2 cells were serum starved overnight (A–C) and amino acid starved 1 h(B and C) before GnRH treatment at indicated doses and time points. Extracts were subjected to SDS-PAGE followed by Western blotting with the following antibodies. A, Anti-4E-BP1 reveals three electrophoretic forms (γ, β α) with the histogram representing the proportion of inactive γ-isoform relative to total 4E-BP1. B, Antiphospho-eIF4E (Ser 209) and eIF4E; histogram represents ratio of phospho-eIF4E relative to total eIF4E. C, Anti-phospho-eIF4G (Ser 1108); histogram represents values expressed as a percentage of maximal induction. Blots are representative images of each experiment. Histograms represent quantitative chemiluminescent image analysis of at least three independent experiments. The asterisks show significant difference from the control mean (P ≤ 0.05) as determined by ANOVA and post hoc Dunnett’s comparison to control test.

Article Snippet: 4E-BP1, phospho-eIF4E, and AKT antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA) whereas phospho-eIF4G, -AKT, -Mnk, and -eIF4E antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: SDS Page, Western Blot, Control, Comparison

Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Acute Regulation of Translation Initiation by Gonadotropin-Releasing Hormone in the Gonadotrope Cell Line LβT2

doi: 10.1210/me.2003-0478

Figure Lengend Snippet: LβT2 cells were serum starved overnight and amino acid starved for 1 h followed with 10 nm GnRH for the times shown. Extracts underwent SDS-PAGE and immunoblotted with the indicated antiserum. A, Three electrophoretic forms (γ, β α) of 4E-BP1; the histogram depicts the proportion of inactive γ-isoform relative to total 4E-BP1. B, Histogram represents proportion of phospho-eIF4E relative to total eIF4E. C, Percent of maximal eIF4G phosphorylation. Blots are representative images. Histograms are the result of quantitative chemiluminescent imaging analysis of at least three separate experiments. The asterisks show significant difference from the control mean (P ≤ 0.05) as determined by ANOVA and post hoc Dunnett’s comparison to control test.

Article Snippet: 4E-BP1, phospho-eIF4E, and AKT antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA) whereas phospho-eIF4G, -AKT, -Mnk, and -eIF4E antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: SDS Page, Phospho-proteomics, Imaging, Control, Comparison

Journal: Molecular endocrinology (Baltimore, Md.)

Article Title: Acute Regulation of Translation Initiation by Gonadotropin-Releasing Hormone in the Gonadotrope Cell Line LβT2

doi: 10.1210/me.2003-0478

Figure Lengend Snippet: LβT2 cells were serum starved overnight, followed by amino acid starvation for 1 h and subsequently pretreated with 10 µm PD98059, 1.0 µm LY294002, or 10 nm rapamycin for 30 min, after which cells were treated with 10 nm GnRH for 15 min. Extracts were separated by SDS-PAGE and immunoblotted with the indicated antiserum. A, Histogram represents activation of phospho-Mnk. B, Histogram represents ratio of phospho-eIF4E to total eIF4E normalized to control. Blots are representative images and histograms are the result of quantitative chemiluminescent imaging analysis of at least three separate experiments. The asterisks show significant difference from the control mean (P ≤ 0.05), as determined by ANOVA and post hoc Dunnett’s comparison to control test.

Article Snippet: 4E-BP1, phospho-eIF4E, and AKT antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA) whereas phospho-eIF4G, -AKT, -Mnk, and -eIF4E antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: SDS Page, Activation Assay, Control, Imaging, Comparison