total p73  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: The p53 R172H Mutant Does Not Enhance Hepatocellular Carcinoma Development and Progression

doi: 10.1371/journal.pone.0123816

Figure Lengend Snippet: A) Immunoblot demonstrating knockdown of p53 in cell lines derived from p53 R172H HCCs as compared to pGIPZ empty vector control infections. B) Quantitative RT-PCR analysis of p53 family transcriptional targets in four mouse HCC cell lines with knockdown of p53 R172H . Ct values in each sample were normalized to β-Actin as an endogenous reference. Fold changes were calculated by normalizing all samples to the average Ct value of the cell line’s pGIPZ control. The Comparative Ct method was used for fold change calculation. C) Immunoblots demonstrating the expression of p63 and p73 isoforms in p53 fl/fl and p53 R172H HCC cell lines. Note the presence of multiple isoforms within the cell lines. GAPDH is used as a loading control. MW = molecular weight marker.

Article Snippet: Antibodies used were: p53 antibody (1:2000, Cell Signaling 1C12), p21 antibody (1:1000, Santa Cruz sc-6246), TAp63 antibody (1:500, Biolegend 618901), ΔNp63 antibody (1:500, Biolegend 619001), Total p63 antibody (1:500 Santa Cruz, ac-8431), TAp73 antibody (1:500 Novus 24737), Total p73 antibody (1:500 Imgenex).

Techniques: Western Blot, Derivative Assay, Plasmid Preparation, Quantitative RT-PCR, Expressing, Molecular Weight, Marker

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type mouse lung microvascular endothelial cells (MLMVEC). Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 5; *P < 0.05, significant interaction by ANOVA). B: wild-type MLMVEC. Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced p73 phosphorylation in wild-type MLMVEC, controlled for total p73 (n = 6; *P < 0.05 by ANOVA). C: protein kinase GI mice (PKGI−/−) MLMVEC. Representative Western blots showing phosphorylation of vasodilator-stimulated phosphoprotein (VASP) serine 235 after 8pCPT-cGMP treatment in wild-type but not PKGI−/− MLMVEC, and the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. D, left: graph shows effect of 30-min 8pCPT-cGMP on H2O2-induced c-Abl activation in PKGI−/− MLMVEC (n = 4; P = not significant). Data are normalized to wild-type mean. D, right: graph shows baseline c-Abl activity, as measured by phospho-Y245 c-Abl/total c-Abl ratio in wild-type vs. PKGI−/− MLMVEC in the absence of 8pCPT-cGMP or H2O2 exposure (n = 9 and 4; *P < 0.01). All values are presented as means ± SE.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay, Activity Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type MLMVEC. Representative western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of atrial natriuretic peptide (ANP; 10 nM). Graph shows effect of 30-min ANP (10 nM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 7; *P < 0.05 by ANOVA). B: wild-type MLMVEC. Representative Western blots showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of ANP. Graph shows effect of H2O2 on p73 phosphorylation, controlled for total p73 (n = 7; *P < 0.05 for effect of H2O2 in diluent-treated cells by ANOVA). The absence of phospho-p73 in ANP-treated cells (30 min, 10 nM) precluded quantification; signal represented as zero in graph.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type mouse lung microvascular endothelial cells (MLMVEC). Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 5; *P < 0.05, significant interaction by ANOVA). B: wild-type MLMVEC. Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced p73 phosphorylation in wild-type MLMVEC, controlled for total p73 (n = 6; *P < 0.05 by ANOVA). C: protein kinase GI mice (PKGI−/−) MLMVEC. Representative Western blots showing phosphorylation of vasodilator-stimulated phosphoprotein (VASP) serine 235 after 8pCPT-cGMP treatment in wild-type but not PKGI−/− MLMVEC, and the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. D, left: graph shows effect of 30-min 8pCPT-cGMP on H2O2-induced c-Abl activation in PKGI−/− MLMVEC (n = 4; P = not significant). Data are normalized to wild-type mean. D, right: graph shows baseline c-Abl activity, as measured by phospho-Y245 c-Abl/total c-Abl ratio in wild-type vs. PKGI−/− MLMVEC in the absence of 8pCPT-cGMP or H2O2 exposure (n = 9 and 4; *P < 0.01). All values are presented as means ± SE.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay, Activity Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type MLMVEC. Representative western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of atrial natriuretic peptide (ANP; 10 nM). Graph shows effect of 30-min ANP (10 nM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 7; *P < 0.05 by ANOVA). B: wild-type MLMVEC. Representative Western blots showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of ANP. Graph shows effect of H2O2 on p73 phosphorylation, controlled for total p73 (n = 7; *P < 0.05 for effect of H2O2 in diluent-treated cells by ANOVA). The absence of phospho-p73 in ANP-treated cells (30 min, 10 nM) precluded quantification; signal represented as zero in graph.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type mouse lung microvascular endothelial cells (MLMVEC). Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 5; *P < 0.05, significant interaction by ANOVA). B: wild-type MLMVEC. Representative Western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of 8pCPT-cGMP. Graph shows effect of 30-min 8pCPT-cGMP (50 μM) on H2O2-induced p73 phosphorylation in wild-type MLMVEC, controlled for total p73 (n = 6; *P < 0.05 by ANOVA). C: protein kinase GI mice (PKGI−/−) MLMVEC. Representative Western blots showing phosphorylation of vasodilator-stimulated phosphoprotein (VASP) serine 235 after 8pCPT-cGMP treatment in wild-type but not PKGI−/− MLMVEC, and the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of 8pCPT-cGMP. D, left: graph shows effect of 30-min 8pCPT-cGMP on H2O2-induced c-Abl activation in PKGI−/− MLMVEC (n = 4; P = not significant). Data are normalized to wild-type mean. D, right: graph shows baseline c-Abl activity, as measured by phospho-Y245 c-Abl/total c-Abl ratio in wild-type vs. PKGI−/− MLMVEC in the absence of 8pCPT-cGMP or H2O2 exposure (n = 9 and 4; *P < 0.01). All values are presented as means ± SE.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay, Activity Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Protein kinase G increases antioxidant function in lung microvascular endothelial cells by inhibiting the c-Abl tyrosine kinase

doi: 10.1152/ajpcell.00375.2012

Figure Lengend Snippet: A: wild-type MLMVEC. Representative western blot showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y245 c-Abl and total c-Abl in the presence and absence of atrial natriuretic peptide (ANP; 10 nM). Graph shows effect of 30-min ANP (10 nM) on H2O2-induced c-Abl activation in wild-type MLMVEC, as measured by c-Abl Y245 phosphorylation, controlled for total c-Abl (n = 7; *P < 0.05 by ANOVA). B: wild-type MLMVEC. Representative Western blots showing the effect of 0, 250, and 500 μM H2O2 on phospho-Y99 p73 and total p73 in the presence and absence of ANP. Graph shows effect of H2O2 on p73 phosphorylation, controlled for total p73 (n = 7; *P < 0.05 for effect of H2O2 in diluent-treated cells by ANOVA). The absence of phospho-p73 in ANP-treated cells (30 min, 10 nM) precluded quantification; signal represented as zero in graph.

Article Snippet: Total p73 and total c-Abl antibodies were from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Western Blot, Activation Assay