Journal: Oncotarget

Article Title: MAGE-A inhibit apoptosis and promote proliferation in multiple myeloma through regulation of BIM and p21 Cip1 .

doi: 10.18632/oncotarget.27488

Figure Lengend Snippet: Figure 2: Silencing of MAGE-A in HMCL results in stabilization of phosphorylated BIM and p21Cip1. MM.1r and H929 were transduced with MAGE-A3-specific lentiviral shRNA constructs or controls as described, and heavy membrane preps (A, C) were probed for expression of Bcl-2 family proteins. (A) Western blot of heavy membrane preps for Bcl-2 family proteins. MAGE-A3 knockdown and increased p53 protein were confirmed by western blot of whole cell lysate. COXIV, load control, representative of all blots. (B) Optical densitometry of western blots for BIM el as in (A) demonstrates significantly higher levels of this protein after MAGE-A silencing compared to controls. Data was pooled from five replicates, representing 13–15 data points each. Error bars, standard error of the mean. (C) Western blot of heavy membrane preps for p-ser69 and p-ser77 Bim el demonstrate stabilization of the phosphorylated protein after MAGE-A knockdown. (D) Whole cell lysates were probed by western blot for p21Cip1. (E) Optical densitometry of western blots for p21Cip1 as in (D) demonstrates significantly higher levels after MAGE-A silencing. Data pooled from five replicates, representing 15 data points each. Con, Untreated control cells. shNT, non-target shRNA lentiviral construct. shMA, MAGE-A3 targeted lentiviral shRNA construct TRCN0000128375.

Article Snippet: The antibodies used were: Bcl-2 (Cell Signaling Technologies, 2872), Bcl-xL (2762), BID (2002), BIM (2819), CoxIV (4850), Mcl-1 (4572), pSer69-BIM (4585), pSer77-BIM (12433), p21 (2947), BMF (Abcam, ab181148), GAPDH (ab8245), Bcl-G (Santa Cruz, sc-393715), Caspase 3 Oncotarget737www.oncotarget.com (sc-7272), MAGE6C1 (sc-20034), PUMA (sc-374223), p53 (sc-126), goat anti-mouse IgG HRP conjugate (Thermo Scientific, 32430) and goat anti-rabbit IgG HRP conjugate (32460).

Techniques: Transduction, shRNA, Construct, Membrane, Expressing, Western Blot, Knockdown, Control

Journal: Oncotarget

Article Title: MAGE-A inhibit apoptosis and promote proliferation in multiple myeloma through regulation of BIM and p21 Cip1 .

doi: 10.18632/oncotarget.27488

Figure Lengend Snippet: Figure 5: Model of MAGE-A3 activity in MM. (A) DNA damage response pathways promote BIM expression and p53 transcriptional activity, resulting in increased expression of BAX and p21Cip1. BIM displaces BAX from anti-apoptotic Bcl-2 family proteins, allowing it to translocate to mitochondrial membranes and initiate apoptosis. P21Cip1 inhibits cyclin D1/CDK4/6 activity, blocking progression through the early G1 checkpoint. (B) MAGE-A3/RING complex activity, possibly activated through interactions with DNA repair pathways, down- regulates BIM and p53 through transcriptional and post-translational mechanisms, resulting in survival and proliferation.

Article Snippet: The antibodies used were: Bcl-2 (Cell Signaling Technologies, 2872), Bcl-xL (2762), BID (2002), BIM (2819), CoxIV (4850), Mcl-1 (4572), pSer69-BIM (4585), pSer77-BIM (12433), p21 (2947), BMF (Abcam, ab181148), GAPDH (ab8245), Bcl-G (Santa Cruz, sc-393715), Caspase 3 Oncotarget737www.oncotarget.com (sc-7272), MAGE6C1 (sc-20034), PUMA (sc-374223), p53 (sc-126), goat anti-mouse IgG HRP conjugate (Thermo Scientific, 32430) and goat anti-rabbit IgG HRP conjugate (32460).

Techniques: Activity Assay, Expressing, Blocking Assay

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Characteristics of patients and analyses of overall survival

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques:

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Overall survival versus Bad and/or Bim phosphorylation. ( a ) Estimation of correlation (Pearson's test) between the level of pBad S136 and pBim S69 . ( b ) Kaplan–Meier curves illustrate the absence of significant difference in OS between patients with tumors harboring low level of pBad S136 and patients with high level of pBad S136 . ( c ) Kaplan–Meier curves illustrate the absence of significant difference in OS between patients with tumors harboring low level of pBim S69 and patients with high level of pBim S69 . ( d ) Kaplan–Meier curves illustrate the significant difference in OS between patients with tumors harboring high levels of pBim S69 and pBad S136 and patients devoid of this double hallmark.

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques:

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Presence of pBad S136 and pBim S69 in GBM and kinase activities. ( a ) Graphs and Pearson's test analyze the presence of correlation between the levels of Akt activity and the level of EGFr activity, or the level of pBad S136 or the level of pBim S69 . ( b ) Graphs and Pearson's test analyze the presence of correlation between the levels of Erk activity and the level of EGFr activity, or the level of pBad S136 or the level of pBim S69 .

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques: Activity Assay