Journal: Molecular medicine reports

Article Title: Caspase-3-resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis.

doi: 10.3892/mmr.2014.3035

Figure Lengend Snippet: Figure 1. Inducible forms of ANP32B expression in the corresponding transformant cells following Tet withdrawal. The (A) U937Tempty, U937TANP32B(WT) and (B) U937TANP32B(D163A) and U937TANP32B(1‑163) cell extracts were detected by western blot analysis for the three forms of ANP32B expression with β‑actin as a loading control. ANP32B, acidic leucine‑rich nuclear phosphoprotein 32B; Tet, tetracycline, WT, wild type.

Article Snippet: The following antibodies were used: Polyclonal rabbit anti-human protein kinase Cδ (PKCδ) (C-20; SC‐937; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); polyclonal rabbit anti-human cleaved caspase-3 (9661; Cell Signaling Technology, Danvers, MA, USA); monoclonal mouse anti-human poly[ADP (adenosine diphosphate)-ribose] polymerase (PARP; F-2; SC-8007; Santa Cruz Biotechnology, Inc.); polyclonal rabbit anti‐human Bcl‐2 (sc‐492; Santa Cruz Biotechnology, Inc.) and polyclonal rabbit anti‐human ANP32B (10843‐1‐AP; ProteinTech Group, Inc., Chicago, IL, USA), with monoclonal mouse anti-β-actin mAb (EMD Millipore, Billerica, MA, USA) to confirm equal loading.

Techniques: Expressing, Western Blot, Control

Journal: Molecular medicine reports

Article Title: Caspase-3-resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis.

doi: 10.3892/mmr.2014.3035

Figure Lengend Snippet: Figure 2. The localization of the different ANP32B forms in the transformant cells following tetracycline withdrawal immunostained with ANP32B antibody in addition to duplicate staining of 4',6‑diamidino‑2‑phenylindole. Images are representative of >20 fields examined. ANP32B, acidic leucine‑rich nuclear phosphoprotein 32B; WT, wild type.

Article Snippet: The following antibodies were used: Polyclonal rabbit anti-human protein kinase Cδ (PKCδ) (C-20; SC‐937; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); polyclonal rabbit anti-human cleaved caspase-3 (9661; Cell Signaling Technology, Danvers, MA, USA); monoclonal mouse anti-human poly[ADP (adenosine diphosphate)-ribose] polymerase (PARP; F-2; SC-8007; Santa Cruz Biotechnology, Inc.); polyclonal rabbit anti‐human Bcl‐2 (sc‐492; Santa Cruz Biotechnology, Inc.) and polyclonal rabbit anti‐human ANP32B (10843‐1‐AP; ProteinTech Group, Inc., Chicago, IL, USA), with monoclonal mouse anti-β-actin mAb (EMD Millipore, Billerica, MA, USA) to confirm equal loading.

Techniques: Staining

Journal: Molecular medicine reports

Article Title: Caspase-3-resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis.

doi: 10.3892/mmr.2014.3035

Figure Lengend Snippet: Figure 3. Uncleavable ANP32B(D163A) protein enhances etoposide‑induced apoptosis. (A) Percentage of annexin‑V+ cells was determined using flow cytom etry, and (B) displayed in a histogram. All values are presented as the mean ± standard deviation; experiments were performed in triplicate. *P<0.05 and **P<0.01 vs. U937Tempty cells. ANP32B, acidic leucine‑rich nuclear phosphoprotein 32B; WT, wild type; PI. propidium iodide.

Article Snippet: The following antibodies were used: Polyclonal rabbit anti-human protein kinase Cδ (PKCδ) (C-20; SC‐937; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); polyclonal rabbit anti-human cleaved caspase-3 (9661; Cell Signaling Technology, Danvers, MA, USA); monoclonal mouse anti-human poly[ADP (adenosine diphosphate)-ribose] polymerase (PARP; F-2; SC-8007; Santa Cruz Biotechnology, Inc.); polyclonal rabbit anti‐human Bcl‐2 (sc‐492; Santa Cruz Biotechnology, Inc.) and polyclonal rabbit anti‐human ANP32B (10843‐1‐AP; ProteinTech Group, Inc., Chicago, IL, USA), with monoclonal mouse anti-β-actin mAb (EMD Millipore, Billerica, MA, USA) to confirm equal loading.

Techniques: Standard Deviation

Journal: Molecular medicine reports

Article Title: Caspase-3-resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis.

doi: 10.3892/mmr.2014.3035

Figure Lengend Snippet: Figure 4. Effect of uncleavable ANP32B(D163A) protein on etoposide‑induced proteolytic cleavage of PKCδ and caspase‑3 activation. (A) Western blot analysis with β‑actin as a loading control. (B) Analysis of caspase‑3 activity. *P<0.05 vs. U937Tempty cells with the corresponding treatment in an independent experi ment with triplicates. ANP32B, acidic leucine‑rich nuclear phosphoprotein 32B; PKC‑δ, protein kinase Cδ; WT, wild type; 1, U937Tempty; 2, U937TANP32B(WT); 3, U937TANP32B(D163A); 4, U937TANP32B(1‑163); PARP, poly[ADP (adenosine diphosphate)‑ribose] polymerase; Con, control; Δ, fragments of the corresponding protein,

Article Snippet: The following antibodies were used: Polyclonal rabbit anti-human protein kinase Cδ (PKCδ) (C-20; SC‐937; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); polyclonal rabbit anti-human cleaved caspase-3 (9661; Cell Signaling Technology, Danvers, MA, USA); monoclonal mouse anti-human poly[ADP (adenosine diphosphate)-ribose] polymerase (PARP; F-2; SC-8007; Santa Cruz Biotechnology, Inc.); polyclonal rabbit anti‐human Bcl‐2 (sc‐492; Santa Cruz Biotechnology, Inc.) and polyclonal rabbit anti‐human ANP32B (10843‐1‐AP; ProteinTech Group, Inc., Chicago, IL, USA), with monoclonal mouse anti-β-actin mAb (EMD Millipore, Billerica, MA, USA) to confirm equal loading.

Techniques: Activation Assay, Western Blot, Control, Activity Assay

Journal: Molecular medicine reports

Article Title: Caspase-3-resistant uncleavable form of acidic leucine-rich nuclear phosphoprotein 32B potentiates leukemic cell apoptosis.

doi: 10.3892/mmr.2014.3035

Figure Lengend Snippet: Figure 5. Uncleavable ANP32B(D163A) induction reduces Bcl‑2 protein expression levels. U937Tempty, U937TANP32B(WT), U937TANP32B(D163A) and U937TANP32B(1‑163) cells were cultured in the absence of tetracycline for different time periods as indicated. Bcl‑2, Mcl‑1 and ICAD proteins were probed by western blot analysis and β‑actin was used as a loading control. ANP32B, acidic leucine‑rich nuclear phosphoprotein 32B; ICAD, inhibitor of caspase‑activated DNase; WT, wild type.

Article Snippet: The following antibodies were used: Polyclonal rabbit anti-human protein kinase Cδ (PKCδ) (C-20; SC‐937; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); polyclonal rabbit anti-human cleaved caspase-3 (9661; Cell Signaling Technology, Danvers, MA, USA); monoclonal mouse anti-human poly[ADP (adenosine diphosphate)-ribose] polymerase (PARP; F-2; SC-8007; Santa Cruz Biotechnology, Inc.); polyclonal rabbit anti‐human Bcl‐2 (sc‐492; Santa Cruz Biotechnology, Inc.) and polyclonal rabbit anti‐human ANP32B (10843‐1‐AP; ProteinTech Group, Inc., Chicago, IL, USA), with monoclonal mouse anti-β-actin mAb (EMD Millipore, Billerica, MA, USA) to confirm equal loading.

Techniques: Expressing, Cell Culture, Western Blot, Control