förster resonance energy transfer fret based akt activity reporter lyn aktar2 ev  (Addgene inc)

Journal: Blood

Article Title: Tonic B-cell receptor signaling in diffuse large B-cell lymphoma

doi: 10.1182/blood-2016-10-747303

Figure Lengend Snippet: Effect of BCR KO on proliferation of GCB-DLBCL lines correlates with reduction in AKT activity. (A) Correlation between relative proliferation after BCR KO in GCB-DLBCL lines, as calculated by the ratio (BCR-KO/BCR replete) of their absolute growth curve slopes, and the BCR KO–induced log2 change in HRK expression. For each cell line, the average value of 2 to 4 biological replicates (each with a different IgH-targeting gRNA) is displayed. (B) Correlation for GCB-DLBCL lines between BCR KO–induced reductions in proliferation and AKT activity, measured by FRET efficiency with the Lyn-AktAR2 reporter. AKT activity reduction value is the average of 3 biological replicates, each with a different IgH-targeting gRNA. (C) Relative decline of KO cells in GCB-DLBCL lines after IgH-targeting KO of BCR (upper panel) or combined KO of all 3 AKT genes (lower panel). Values shown are the mean ± SD from 2 (BCR KO) or 3 (AKT KO) biological replicates. (D) Relative decline of BCR-KO cells in DLBCL lines that are either unmodified or have previously undergone PTEN KO. Values shown are the mean ± SD from 3 biological replicates.

Article Snippet: Förster resonance energy transfer (FRET) The FRET-based AKT activity reporter AktAR2 (Addgene) 12 was modified by N-terminal addition of the 10 N-terminal amino acids (AA) from Lyn kinase.

Techniques: Activity Assay, Expressing