Journal: Cell Transplantation

Article Title: Impact of One Versus Two Consecutive Doses of Endothelial Cells (EPCs) and EPCs-Derived Condition Medium on Protecting Myocardium from Acute Ischemia-Reperfusion Injury in Rat

doi: 10.1177/09636897211007049

Figure Lengend Snippet: Protein expressions of apoptotic, fibrotic, mitochondrial-damaged, and pressure overload/heart failed biomarkers in LV myocardium by day 3 after IR induction. (A) Protein expressions of cleaved caspase 3 (c-Csp3), * vs. other groups with different symbols (†, ‡, §, ¶), P < .0001. (B) Protein expression of cleaved poly (ADP-ribose) polymerase (c-PARP), * vs. other groups with different symbols (†, ‡, §, ¶), P < .0001. (C) Protein expression of Smad3, * vs. other groups with different symbols (†, ‡, §, ¶, λ), P < .0001. (D) Protein expression of transforming growth factor (TGF)-ß, * vs. other groups with different symbols (†, ‡, §, ¶), P < .0001. (E) Protein expression of cytosolic cytochrome C (cyto-CytC), * vs. other groups with different symbols (†, ‡, §, ¶), P < .0001. (F) Protein expression of brain natriuretic peptide (BNP) * vs. other groups with different symbols (†, ‡, §, ¶), P < .0001. All statistical analyses were performed by one-way ANOVA, followed by Bonferroni multiple comparison post hoc test ( n = 6 for each group). Symbols (*, †, ‡, §, ¶) indicate significance (at .05 level). Group 1 = sham-operated control; group 2 = ischemia-reperfusion (IR) + culture medium; group 3 = IR + endothelial progenitor cells (EPCs) administered at 3 h after IR induction; group 4 = IR + EPCs administered at 3 h and 24 h after; group 5 = IR + condition medium (CM) administered at 3 h after IR procedure; group 6 = IR + CM administered 3 h and 24 h after IR procedure.

Article Snippet: The membranes were incubated with the indicated primary antibodies [CD31 (1:1000, Abcam, Cambridge, UK), von Willebrand factor (vWF) (1:1000, Abcam, Cambridge, UK), endothelial nitric oxide synthase (eNOS) (1:1000, Abcam, Cambridge, UK), cleaved poly (ADP-ribose) polymerase (PARP) (1:1000, Cell Signaling, Danvers, Ma, USA), cleaved caspase 3 (c-Csp3) (1: 1000, Cell Signaling, Danvers, Ma, USA), Smad3 (1:1000, Cell Signaling, Danvers, Ma, USA), brain natriuretic peptide (BNP) (1:500, Abcam, Cambridge, UK), NADPH oxidase (NOX)-1 (1:1500, Sigma, St. Louis, MO, USA), NOX-2 (1:500, Sigma, St. Louis, MO, USA), cytosolic cytochrome C (cyto-CytC) (1:2000, BD, Franklin Lakes, NJ, USA), mitochondrial cytochrome C (Mito CytC) (1:2000, BD, Franklin Lakes, NJ, USA), p47phox (1:1000, Merck Millipore, Burlington, Ma, USA), transforming growth factor (TGF)-ß (1:5000, Abcam), vascular endothelial growth factor (VEGF) (1:1000, Abcam, Cambridge, UK), CXCR4 (1:1000, Abcam, Cambridge, UK), stromal cell-derived factor (SDF-1α) (1:1000, Cell Signaling, Danvers, Ma, USA) and hypoxia-inducible factor (HIF)-1α (1:1000, Abcam, Cambridge, UK) for 1 hour at room temperature.

Techniques: Expressing

Journal:

Article Title: Biochemical and Genetic Analysis of the Mitochondrial Response of Yeast to BAX and BCL-X L

doi:

Figure Lengend Snippet: BAX localizes to mitochondria but does not cause detectable release of cyt c. (A) Subcellular distribution of BAX in yeast cells. Cells transformed with GAL vector (lanes 1 to 4) or with GAL-BAX (lanes 5 to 8) were suspended in isotonic buffer, homogenized, and separated into soluble fraction (S), light-membrane fraction (LM), heavy-membrane fraction (HM), and low-speed pellet (P1) by differential centrifugation. The fractions were analyzed by Western blotting using anti-mBAX Ab 651 (top panel), anti-yVDAC1 Ab (middle panel), and anti-cyt c MAb (PharMingen) (bottom panel). The P1 pellet contains residual whole cells, nuclei, and mitochondria. The HM fraction is enriched for intact mitochondria. The LM fraction contains the endoplasmic reticulum, plasma membrane, and mitochondrial fragments. The soluble (S) fraction represents the cytosol. ×3 HM denotes that three times the amount of protein loaded in lane 7 was loaded in this lane. (B) BAX is enriched in mitochondrial contact sites. Submitochondrial membrane vesicles were prepared from isolated yeast mitochondria expressing HA-BAX by sonication and separated by centrifugation through a continuous sucrose density gradient. Fractions were analyzed by Western blotting using anti-yF1β-ATPase Ab, anti-yVDAC1 Ab, anti-yANT Ab, and anti-mBAX Ab 651. The high-density fractions (HDF) are enriched in inner membrane (IM) vesicles. The low-density fractions (LDF) are enriched in outer mitochondrial membranes (OM). Intermediate-density fractions (IDF) are enriched for contact sites (CS), sites of association between the inner and outer membranes.

Article Snippet: The fractions were analyzed by Western blotting using anti-mBAX Ab 651 (top panel), anti-yVDAC1 Ab (middle panel), and anti-cyt c MAb (PharMingen) (bottom panel).

Techniques: Transformation Assay, Plasmid Preparation, Centrifugation, Western Blot, Isolation, Expressing, Sonication