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(A) Western blot of total 4E-BP in RPE-1 asynchronous, mitotic-enriched, and G2-synchronized cells. (B) Western blot of non-phosphorylated, phosphorylated Threonine 70, and phosphorylated Serine 63 4E-BP. (C) Cap-pulldown analysis with <t>m</t> <t>7</t> GTP agarose beads with asynchronous, mitotic-enriched, and G2-synchronized RPE-1 cells. Western blot of 4E-BP total and eIF4E1. (D) Western blot analysis of phosphorylated and total Rps6 in RPE-1 asynchronous cells, mitotic-enriched cells, and treated with an mTOR inhibitor (INK-128). (E) Quantification of normalized phosphorylated to total Rps6. Significance determined by one-way ANOVA with Dunnett’s post-hoc, asynchronous reference. Error bars represent the standard deviation, n=3 biological replicates
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(A) Western blot of total 4E-BP in RPE-1 asynchronous, mitotic-enriched, and G2-synchronized cells. (B) Western blot of non-phosphorylated, phosphorylated Threonine 70, and phosphorylated Serine 63 4E-BP. (C) Cap-pulldown analysis with m 7 GTP agarose beads with asynchronous, mitotic-enriched, and G2-synchronized RPE-1 cells. Western blot of 4E-BP total and eIF4E1. (D) Western blot analysis of phosphorylated and total Rps6 in RPE-1 asynchronous cells, mitotic-enriched cells, and treated with an mTOR inhibitor (INK-128). (E) Quantification of normalized phosphorylated to total Rps6. Significance determined by one-way ANOVA with Dunnett’s post-hoc, asynchronous reference. Error bars represent the standard deviation, n=3 biological replicates

Journal: bioRxiv

Article Title: Decreased tRNA abundance contributes to decreased translation elongation rate in a prolonged mitosis

doi: 10.64898/2026.01.13.699274

Figure Lengend Snippet: (A) Western blot of total 4E-BP in RPE-1 asynchronous, mitotic-enriched, and G2-synchronized cells. (B) Western blot of non-phosphorylated, phosphorylated Threonine 70, and phosphorylated Serine 63 4E-BP. (C) Cap-pulldown analysis with m 7 GTP agarose beads with asynchronous, mitotic-enriched, and G2-synchronized RPE-1 cells. Western blot of 4E-BP total and eIF4E1. (D) Western blot analysis of phosphorylated and total Rps6 in RPE-1 asynchronous cells, mitotic-enriched cells, and treated with an mTOR inhibitor (INK-128). (E) Quantification of normalized phosphorylated to total Rps6. Significance determined by one-way ANOVA with Dunnett’s post-hoc, asynchronous reference. Error bars represent the standard deviation, n=3 biological replicates

Article Snippet: Jena Bioscience m 7 GTP-sepharose beads (NU-1122) were incubated with 600-1000 μg of protein and tumbled for one hour.

Techniques: Western Blot, Standard Deviation