Journal: Journal of the American Heart Association

Article Title: Impaired Cardiac AMPK (5′‐Adenosine Monophosphate‐Activated Protein Kinase) and Ca ²⁺ ‐Handling, and Action Potential Duration Heterogeneity in Ibrutinib‐Induced Ventricular Arrhythmia Vulnerability

doi: 10.1161/jaha.123.032357

Figure Lengend Snippet: Figure 6. Effect of PI3K-Akt pathway, CaMKII, CSK, and AMPK activity after 4 weeks of oral gavage. A, Western blot (left) and densitometric analysis for p-AMKP, AMPK, CSK, and p-AMKP/AMPK in control and ibrutinib- treated rats’ hearts. Graphs are mean±SEM. n=8 rats in each group. *P<0.05; **P<0.01. The relative protein expression levels of p-AMPK, AMPK, and p-AMPK/AMPK in the control and ibrutinib groups were compared by unpaired t test. The relative protein expression levels of CSK in the control and ibrutinib groups were compared by Mann–Whitney test. B, Western blot (left) and densitometric analysis for ox-CaMKII, p-CaMKII, and CaMKII in control and ibrutinib-treated rats’ hearts. Graphs are mean±SEM. n=8 rats in each group. The relative protein expression levels of ox-CaMKII, p-CaMKII, and CaMKII in the control and ibrutinib groups were compared by unpaired t test. C, Western blot (left) and densitometric analysis for PI3K, p- Akt, and Akt in control and ibrutinib-treated rats’ hearts. Graphs are mean±SEM. n=8 rats in each group. The relative protein expression levels of PI3K and p-Akt in the control and ibrutinib groups were compared by an unpaired t test. The relative protein expression levels of Akt in the control and ibrutinib groups were compared by Mann–Whitney test. Akt indicates protein kinase B; AMPK, 5′-adenosine monophosphate-activated protein kinase; CaMKII, calcium/calmodulin-dependent kinase, type II; CSK, C-terminal src kinase; Ctrl, control group; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; Ibr, ibrutinib group; ns, not significant; ox-CaMKII, oxidized CaMKII; p-Akt, phosphorylated Akt; p-AMPK, phosphorylated AMPK; p-CaMKII, phosphorylated CaMKII; and PI3K, phosphoinositide 3′-kinase.

Article Snippet: To prevent nonspecific protein binding, the membranes were preincubated with 5% bovine serum albumin before the incubation overnight with specific antibodies against total and phosphoPLB (Badrilla), SERCA2a (Badrilla), Cav1.2- pSer1928 (Badrilla), Cav1.2 (Sigma), NCX (biorbyt), RyR2 (Thermo Scientific), RyR2- pSer2814, RyR2- pSer2808 (Badrilla), PI3K110α (Cell Signaling Technology), total and phospho- Akt (Cell Signaling Technology), total and phospho- AMPK (Cell Signaling Technology), CSK (Cell Signaling Technology), ox- CaMKII (Met281/282) (Sigma), CaMKII (pThr286 or 287) (Badrilla), CaMKIIδ (abcam), and GAPDH (Fitzgerald) overnight.

Techniques: Activity Assay, Western Blot, Control, Expressing, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Systemic lupus erythematosus immune complexes increase the expression of SLAM family members CD319 (CRACC) and CD229 (LY-9) on plasmacytoid dendritic cells and CD319 on CD56(dim) NK cells.

doi: 10.4049/jimmunol.1301022

Figure Lengend Snippet: FIGURE 5. pDCs lack the activating adaptor molecules but express inhibitory adaptor molecules of SLAM receptors. (A) IFN-a production by RNA-IC–stimulated pDCs after cross-linking of CD319, CD229, or BDCA-2 by plate-bound mAbs. IFN-a levels were determined in 20-h cell culture supernatants by an immunoassay and normalized to the IFN-a production in cultures without plate-bound mAb. The dashed line indicates 100%. Data (mean + SD) are from three donors from two independent experiments. Expression of EAT2 and SAP (B) and SHIP-1, SHP-1, SHP- 2, and CSK (C) in cell lysates from isolated pDCs or NK cells from two healthy donors was determined by Western blot. Expression of b-actin was used as control.

Article Snippet: Rabbit polyclonal Abs to Ewing’s sarcoma-activated transcript 2 (EAT2), SHIP-1, SHP-2, CSK (Proteintech Group, Chicago, IL), and a rabbit mAb to SHP-1 (EPR5519; Epitomics, Burlingame, CA), followed by HRP-conjugated goat anti-rabbit IgG (H+L) (Invitrogen), were used to detect the indicated signaling molecules.

Techniques: Cell Culture, Expressing, Isolation, Western Blot, Control