livin antibody  (Bio-Techne corporation)

Journal: Immunity, inflammation and disease

Article Title: Dendritic cells infected with recombinant adenoviral vector encoding mouse fibroblast activation protein-α and human livin α exert an antitumor effect against Lewis lung carcinoma in mice.

doi: 10.1002/iid3.1011

Figure Lengend Snippet: FIGURE 1 Identification of rAd‐infected DCs in vitro. (A–C) rAd‐FAP, rAd‐hlivin α, and rAd‐FAP/hlivin α were constructed using mouse FAP cDNA and human livin α cDNA, and then transduced into mouse bone marrow‐derived DCs, respectively. For evaluation of transduction efficacy, western blot was performed to measure protein expressions of FAP and livin α in cells, with GAPDH functioned as a loading control. ***p < .001, versus 1#; +++p < .001, versus 2#; ^^^p < .001, versus 3#. DCs, dendritic cells; FAP, fibroblast activation protein‐α; rAd‐FAP, recombinant adenoviral vector encoding mouse FAP cDNA; rAd‐hlivin α, recombinant adenoviral vector encoding human livin α cDNA.

Article Snippet: Following incubation with blocking buffer (P0231, Beyotime), Western blot was performed by incubating the membranes with primary antibodies against FAP (NB110‐85534, 88 kDa, Novus Biologicals), livin α (NB100‐56145, 33 kDa, Novus Biologicals), CD45 (ab10558, 147 kDa, Abcam), CD31 (ab281583, 82 kDa, Abcam), α‐SMA (ab5694, 42 kDa, Abcam), and internal control GAPDH (ab8245, 37 kDa, Abcam) at 4°C overnight.

Techniques: Infection, In Vitro, Construct, Derivative Assay, Transduction, Western Blot, Control, Activation Assay, Recombinant, Plasmid Preparation

Journal: Immunity, inflammation and disease

Article Title: Dendritic cells infected with recombinant adenoviral vector encoding mouse fibroblast activation protein-α and human livin α exert an antitumor effect against Lewis lung carcinoma in mice.

doi: 10.1002/iid3.1011

Figure Lengend Snippet: FIGURE 2 Identification of LLC tumor‐isolated CAFs and determination on expressions of FAP and livin α in LLC. (A–C) Western blot was performed to measure protein expressions of CD45, CD31, α‐SMA, and FAP in CAFs isolated from LLC‐bearing mice as well as the protein expression of livin α in mouse LLC tumors. GAPDH was used as the loading control. +++p < .001, versus lysate; ^^^p < .001, versus tumor; ##p < .001, versus Para. CAFs, cancer‐associated fibroblasts; LLC, Lewis lung carcinoma; Para, paracancerous tissues.

Article Snippet: Following incubation with blocking buffer (P0231, Beyotime), Western blot was performed by incubating the membranes with primary antibodies against FAP (NB110‐85534, 88 kDa, Novus Biologicals), livin α (NB100‐56145, 33 kDa, Novus Biologicals), CD45 (ab10558, 147 kDa, Abcam), CD31 (ab281583, 82 kDa, Abcam), α‐SMA (ab5694, 42 kDa, Abcam), and internal control GAPDH (ab8245, 37 kDa, Abcam) at 4°C overnight.

Techniques: Isolation, Western Blot, Expressing, Control

Journal: Immunity, inflammation and disease

Article Title: Dendritic cells infected with recombinant adenoviral vector encoding mouse fibroblast activation protein-α and human livin α exert an antitumor effect against Lewis lung carcinoma in mice.

doi: 10.1002/iid3.1011

Figure Lengend Snippet: FIGURE 3 The antitumor effect of rAd‐FAP/hlivin α‐transduced DCs on LLC in mice. (A and B) Mice were separately immunized with rAd‐EGFP‐transduced DCs, rAd‐FAP‐transduced DCs, rAd‐hlivin α‐transduced DCs, and rAd‐FAP/hlivin α‐transduced DCs (5 × 105 cells/ mouse) on Day 8 postinjection with LLC cells, and tumor volume (mm3) was calculated every 2 days for a total 24 days. (C) Mouse survival rate was observed for 80 days. (D) Splenic lymphocytes were obtained from mice on Day 7 after receiving injection with rAd‐EGFP‐ transduced DCs, rAd‐FAP‐transduced DCs, rAd‐hlivin α‐transduced DCs or rAd‐FAP/hlivin α‐transduced DCs. The cytotoxic effect of splenic lymphocytes on LLC‐derived CAFs was analyzed using Cytotox96 Non‐Radioactive Cytotoxicity Assay Kit. ***p < .001, versus 1#; +p < .05, +++p < .001, versus 2#; ^p < .05, ^^p < .01, ^^^p < .001, versus 3#. CAFs, cancer‐associated fibroblasts; DCs, dendritic cells; LLC, Lewis lung carcinoma; FAP, fibroblast activation protein‐α; rAd‐FAP, recombinant adenoviral vector encoding mouse FAP cDNA; rAd‐hlivin α, recombinant adenoviral vector encoding human livin α cDNA.

Article Snippet: Following incubation with blocking buffer (P0231, Beyotime), Western blot was performed by incubating the membranes with primary antibodies against FAP (NB110‐85534, 88 kDa, Novus Biologicals), livin α (NB100‐56145, 33 kDa, Novus Biologicals), CD45 (ab10558, 147 kDa, Abcam), CD31 (ab281583, 82 kDa, Abcam), α‐SMA (ab5694, 42 kDa, Abcam), and internal control GAPDH (ab8245, 37 kDa, Abcam) at 4°C overnight.

Techniques: Injection, Derivative Assay, Cytotoxicity Assay, Activation Assay, Recombinant, Plasmid Preparation

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Relative livin mRNA expression levels in normal adrenal glands (NAG), adrenocortical adenoma (ACA) and adrenocortical carcinoma (ACC) in the entire cohort of patients, expressed in log scale ( A ) and in 19 paired samples of tumor and corresponding NAG, of which 6 ACC ( B ) and 13 ACA ( C ) ( p < 0.005 per trend). Statistical analysis by Kruskall-Wallis test. N.s.= p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Expressing

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) 4× agarose gel for the expression of livin isoforms α and β at mRNA analyzed by RT-PCR in a subgroup of 4 paired samples of tumors (2 adrenocortical adenomas = ACA, and 2 adrenocortical carcinomas = ACC) and adjacent normal = NAG. Hela cells were used as positive control and β2-microglobulin was used as internal standard. ( B ) Quantitative analysis of agarose gel bands of all 15 paired samples of adrenocortical tumors (ACT) and corresponding NAG ( p = 0.3 per trend). Each bar in the histograms represents the mean of the ratio livin α or livin β signal to β2-microglobulin signal. ( C ) Western blot analysis of livin isoforms expression in three of four paired samples of tumors (2 ACA and 1 ACC) and adjacent NAG that are showed also at mRNA levels. Whole cell lysate SK-MEL 28 was used as positive control and β-actin was used as internal standard. ( D ) Quantitative analysis of WB bands of all 15 paired samples of tumors and corresponding NAG ( p < 0.0001 per trend). Each bar in the histograms represents the mean of the ratio livin α or livin β signal to β-actin signal. Statistical analysis by Kruskall-Wallis test and Mann-Whitney U test. N.s. = p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Agarose Gel Electrophoresis, Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Western Blot, MANN-WHITNEY

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) Normal adrenal gland (NAG) (A.1–2) with adjacent adrenocortical carcinoma (ACC) (A 3-4) stained for H&E (A.1 and A.3) and livin (A.2 and A.4). A.1b, 2b and A.3b, 4b are 10× enlarged and detailed images of the 2× NAG and ACC, respectively. Arrows point to tumor infiltration in the adjacent NAG. TC: capsule, separating NAG and ACC. ( B ) Example of ACC with H-score 3 and negative nuclei stained for H&E (B.1) and livin (B.2) in comparison with ( C ) adrenocortical adenoma (ACA) characterized by H-score 2 and positive nuclei stained for H&E (C.1) and livin (C.2). The .1b, 2b and C.1b, C.1c are 20× enlarged and detailed images of the 10× ACC and ACA, respectively. Scale bar: 100 μm.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Staining, Comparison

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Livin staining in normal adrenal gland ( A ). It is possible to recognize the three different zone of the adrenal gland: the zona glomerulosa (ZG), the zona fasciculata (ZF) and the zona reticularis (ZR). The Figures ( B ) and ( C ) are 20× enlarged and detailed images of the 10× ZG, ZF and ZR, respectively. Scale bar: 100 μm.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Staining

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: ( A ) Livin cytoplasmic protein expression evaluated as H-score in normal adrenal glands (NAG, n = 20), adrenocortical adenomas (ACA, n = 58) and adrenocortical carcinomas (ACC, n = 192) ( p = 0.01 per trend). ( B ) Livin cytoplasm/nuclear protein expression ratio evaluated as the ratio between H-score and nuclear score in NAG ( n = 20), ACA ( n = 58) and ACC ( n = 192) ( p < 0.0001 per trend). Statistical analysis by Kruskall-Wallis test. N.s. = p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Expressing

Journal: Oncotarget

Article Title: Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

doi: 10.18632/oncotarget.14067

Figure Lengend Snippet: Relative livin mRNA levels ( A ) and CASP3 mRNA levels ( B ) evaluated by qRT-PCR at both 48 h and 72 h in NCI-H295R cells after livin transfection in comparison with those transfected with the empty vector (pCMV6). All qRT-PCR experiments were conducted in triplicates. (A) After 72 hours from transfection, the mRNA level of livin α and livin β were significantly increased in livin -NCI-H295R cells in comparison with those transfected with empty vector ( p = 0.04 and p = 0.02 for livin α and livin β , respectively). (B) Livin overexpression was associated with a significant decrease of CASP3 mRNA levels after 72 hours from transfection ( p = 0.015 ). ( C ) WB analysis showed clearly higher livin α and β protein levels and a light decrease of cleaved caspase 3 protein levels at both 48 and 72 hours. ( D ) Quantitative analysis of livin α, livin β and cleaved caspase-3 by WB in livin -NCI- H295R transfected cells in comparison with those transfected with the empty vector after 72 h. Livin α and livin β were significantly increased in livin -NCI-H295R cells in comparison with pCMV6-NCI-H295R ( p = 0.0028 and p = 0.008 for livin α and livin β, respectively). Cleaved caspase-3 was only slightly decreased in livin -NCI- H295R transfected cells ( p = n.s .). Each bar in the histograms represents the mean of the ratio between cleaved casp3 signal and α-tubulin signal. Each point represents the value of this ratio deriving from four separate experiments. ( E ) Immunofluorescence staining showed a decrease of cleaved caspase-3 protein levels (red color) in livin -NCI-H295R cells (green color) compared to pCMV6-NCI-H295R. Cell nuclei were visualized with 4#-6-diamidino-2-phenylindole (DAPI, blue color). Magnification 60×. Statistical analysis by unpaired t-test with Welch's correction. n.s.= p not significant.

Article Snippet: Tissue sections were incubated at room temperature for 1 hour with the primary polyclonal rabbit anti-livin Ab (NB100-56145, Novus Biologicals, 1:1000).

Techniques: Quantitative RT-PCR, Transfection, Comparison, Plasmid Preparation, Over Expression, Immunofluorescence, Staining

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 1. Livin mRNA and protein expression levels in tissue specimens. (A) Semi-quantative RT-PCR values for livin mRNA level in 50 EOC, 20 benign ovarian tumors and 20 normal ovarian tissues. Bar represents the mean value (1.0) for livin expression in positive primary tumors. (B) Representative expression of livin mRNA detected by RT-PCR for positive specimens. (C) Representative livin protein expression and β-actin (internal control) by western blot analysis for cor responding specimens. The data were normalized to the internal control β-actin, and represent the mean values ± SD.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 2. The livin shRNA lentivirus silenced livin expression in the SKOV3 cells. Effective livin targets that simultaneously aimed at both of the two livin iso mers in the livin gene were designed, synthesized, and packaged by lentiviruses. (A) SKOV3 cells were successfully infected with livin Psi-4 shRNA lentivirus at MOI 20 for 72 h; most of the survived cells were GFP positive (original magnification, x400). (B) The livin mRNA expression decreased significantly in 72 h post-infection with livin Psi-4 shRNA detected by real-time RT-PCR. The data were normalized to the untreated control. (C) Immunoblotting evaluations showed that livin protein expression was significantly reduced at 72 h post-infection. The data were normalized to the internal control β-actin, and represent the mean values ± SD. *p<0.05 vs. untreated.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: shRNA, Expressing, Synthesized, Infection, Quantitative RT-PCR, Control, Western Blot

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 3. Silencing of livin gene expression by RNAi induced cell apoptosis in SKOV3 cells. (A) MTT assay showed that the cell proliferation of SKOV3 cells infected with livin shRNA was significantly decreased compared with the controls. (B and C) Flow cytometry revealed that the percentage of apoptotic cells (determined by Annexin V-positive and 7-AAD-negative cells) were increased in the livin-siRNA infected cells, compared to the controls. *p<0.05 vs. untreated.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: Gene Expression, MTT Assay, Infection, shRNA, Flow Cytometry

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 4. Impact of livin lentivirus vectors on cleavage of caspase 9, 7 and 3 expression levels. Western blot analysis showed cleaved caspase 9, 7 and 3 protein levels were remarkably increased, after livin shRNA lentivirus infection of SKOV3 cells. The data were normalized to the internal control β-actin, and represent the mean values ± SD. *p<0.05 vs. untreated.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: Expressing, Western Blot, shRNA, Infection, Control

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 5. Inhibition of livin expression sensitizes SKOV3 cells toward DPP stimulus. (A) Inhibition curves of DPP at seven different test drug concentrations. SKOV3 cell knockdown of livin showed higher sensitivity to DPP as evidenced by lower IC50. (B) Flow cytometry revealed that DPP-induced apoptotic ratio in SKOV3 cells was notably increased after silencing of the livin. *p<0.05 vs. untreated.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: Inhibition, Expressing, Knockdown, Flow Cytometry

Journal: International journal of oncology

Article Title: Expression and role of the inhibitor of apoptosis protein livin in chemotherapy sensitivity of ovarian carcinoma.

doi: 10.3892/ijo.2012.1540

Figure Lengend Snippet: Figure 6. DPP-induced apoptosis in SKOV3 cells before and after silencing of the livin. DPP activated caspase 9, 7 and 3 protein levels were detected by immunoblotting in SKOV3 cells with or without silencing the livin. Inhibition of livin exhibited synergistic effect on induction of apoptosis, with remarkable enhancement of caspase 9, 7 and 3 cleavages compared with the controls. The data were normalized to the internal control β-actin, and represent the mean values ± SD. *p<0.05 vs. untreated, #p<0.05 vs. DPP.

Article Snippet: The membrane was then blocked with 5% nonfat milk and immunoprobed with rabbit polyclonal anti-livin antibody (IMG-347, Imgenex, San Diego, CA, USA), followed by exposed to horseradish peroxidase-conjugated secondary antibody and visualized using ECL plus reagent (GE Healthcare, Piscataway, NJ, USA).

Techniques: Western Blot, Inhibition, Control