Journal: Cell Death & Disease

Article Title: GNA13 regulates BCL2 expression and the sensitivity of GCB-DLBCL cells to BCL2 inhibitors in a palmitoylation-dependent manner

doi: 10.1038/s41419-020-03311-1

Figure Lengend Snippet: A , B Relationship between survival of GCB-DLBCL and expression levels of GNA13 and BCL-2 was analyzed based on public datasets (PPISURV). ( A ) High-level expression of GNA13 is significantly associated with positive outcome of GCB-DLBCL. ( B ) Overexpression of BCL-2 is significantly associated with negative outcome of GCB-DLBCL. C Plot of GNA13 expression vs. BCL2 expression in 102 GCB-DLBCL cases (R2: Genomics Analysis and Visualization Platform, https://hgserver1.amc.nl/cgi-bin/r2/main.cgi ). The transcript levels of GNA13 and BCL2 are inversely related ( r = −0.729, P = 7.155 × 10 −6 ). D Western blot analysis of GNA13 and BCL2 expressions in SU-DHL4 cells, which harbor the WT GNA13, OCI-LY1, and SU-DHL6 cells, both of which harbor mutant GNA13. α-Tubulin was used as a loading control. E Western blot analysis of GNA13 and BCL2 expressions in SU-DHL4 cells expressing scrambled shRNA (Scr) or GNA13 knockdown shRNA (UTR) (left two lanes), and the SU-DHL4-shGNA13 UTR cells ectopically overexpressing (OE) HA-tagged WT, C14S, C18S, or C14/18S mutant GNA13 (right four lanes). GAPDH was used as a loading control. F Western blot analysis of GNA13 and BCL2 expressions in OCI-LY1 cells expressing scrambled shRNA (Scr) or GNA13 knockdown shRNA (UTR) (left two lanes) and the OCI-LY1 cells ectopically expressing HA-tagged WT, C14S, C18S, or C14/18S mutant GNA13 (right four lanes). GAPDH was used as a loading control.

Article Snippet: The BCL2 inhibitor venetoclax (ABT-199), the pan-caspase inhibitor Emricasan and Q-VD-Oph, the group II caspases inhibitor Ac-DEVD-CHO, the pan-PI3K inhibitor GDC-0941, and the PI3Kα/δ inhibitor Copanlisib were purchased from CSNpharm (Arlington Heights, IL, USA).

Techniques: Expressing, Over Expression, Western Blot, Mutagenesis, Control, shRNA, Knockdown

Journal: Cell Death & Disease

Article Title: GNA13 regulates BCL2 expression and the sensitivity of GCB-DLBCL cells to BCL2 inhibitors in a palmitoylation-dependent manner

doi: 10.1038/s41419-020-03311-1

Figure Lengend Snippet: A Volcano plot of FDA-approved drugs and bioactive compounds with known targets on SU-DHL4-shGNA13 Scr and SU-DHL4-shGNA13 UTR cells. Drug effect size ratio between SU-DHL4-shGNA13 Scr and SU-DHL4-shGNA13 UTR vs. statistical significance ( P value) were plotted. Red and blue points indicate drug identified as differentially inhibited between the two types of cells. B Dose–response curves for SU-DHL4-shGNA13 Scr or SU-DHL4-shGNA13 UTR treated with AKT inhibitor MK-2206. C IC 50 of BCL2 inhibitors ABT-199 and ABT-263 in DLBCL cell lines with either wild-type (blue dots) or mutant (red dots) GNA13 . D Dose–response curves for SU-DHL4 and OCI-LY1 cells treated with ABT-199. E SU-DHL4 or OCI-LY1 cells were transfected with constructs containing shGNA13-600, shGNA13-UTR, or a scrambled shRNA (Scr). Levels of total (T)-AKT and phosphorylated (P)-AKT S473 in these cells were examined by western blot analysis. GAPDH was used as loading controls. F Western blot analysis of BCL2, total (T)-AKT, and phosphorylated (P)-AKT S473 level in SU-DHL4 cells treated with PI3K inhibitors Copanlisib or GDC-0941 at indicated concentrations for 6 h, respectively. G Western blot analysis of BCL2 level in SU-DHL4-shGNA13 Scr or SU-DHL4-shGNA13 UTR cells treated with PI3K inhibitors Copanlisib or GDC-0941 at indicated concentrations for 6 h, respectively. α-Tubulin was used as a loading control.

Article Snippet: The BCL2 inhibitor venetoclax (ABT-199), the pan-caspase inhibitor Emricasan and Q-VD-Oph, the group II caspases inhibitor Ac-DEVD-CHO, the pan-PI3K inhibitor GDC-0941, and the PI3Kα/δ inhibitor Copanlisib were purchased from CSNpharm (Arlington Heights, IL, USA).

Techniques: Mutagenesis, Transfection, Construct, shRNA, Western Blot, Control

Journal: Cell Death & Disease

Article Title: GNA13 regulates BCL2 expression and the sensitivity of GCB-DLBCL cells to BCL2 inhibitors in a palmitoylation-dependent manner

doi: 10.1038/s41419-020-03311-1

Figure Lengend Snippet: A Dose–response curves for SU-DHL4-shGNA13 Scr and SU-DHL4-shGNA13 UTR cells treated with ABT-199. B Tumor growth curves of SU-DHL4-shGNA13 UTR cells either expressing the WT GNA13 (SU-DHL4-shGNA13 UTR -OE WT ) or the C14/C18S palmitoylation mutant of GNA13 (SU-DHL4-shGNA13 UTR -OE C14/18S ) in mice treated with either BCL2 inhibitor ABT-199 or vehicle. n , number.

Article Snippet: The BCL2 inhibitor venetoclax (ABT-199), the pan-caspase inhibitor Emricasan and Q-VD-Oph, the group II caspases inhibitor Ac-DEVD-CHO, the pan-PI3K inhibitor GDC-0941, and the PI3Kα/δ inhibitor Copanlisib were purchased from CSNpharm (Arlington Heights, IL, USA).

Techniques: Expressing, Mutagenesis