Supplementary Data - PDF file 3276K, Supplementary Figure 1. Characterization of MMTV-PPAR{delta}... more Supplementary Data - PDF file 3276K, Supplementary Figure 1. Characterization of MMTV-PPAR{delta} transgenic mice. Supplementary Figure 2. Metabolomic analysis Supplementary Figure 3. Signaling pathways associated with PPRE-containing genes. activated in MMTV-PPAR{delta} mice treated with GW501516 for 11 weeks. Supplementary Table 1. Antibodies for IHC and western blotting. Supplementary Table 2. List of primers for qRT-PCR analysis. Supplementary Table 3. Gene expression preferentially altered in MMTV-PPAR{delta} mice treated with GW501516 and everolimus. Supplementary Table 4. Gene expression preferentially altered in MMTV-PPARd mice treated with GW501516 for 11 weeks. Supplementary Table 5. Gene expression preferentially altered in MMTV-PPARd mice. Supplementary Table 6. Gene expression preferentially altered in wild-type mice treated with GW501516 for 11 weeks.
In previous studies we have found that intact AtT-20 cells contained two glucocorticoid binding s... more In previous studies we have found that intact AtT-20 cells contained two glucocorticoid binding sites with distinctly different affinities and specificity. In this paper, the nature of these sites was investigated by studying glucocorticoid binding to cytosol and to plasma membranes isolated from AtT-20 mouse pituitary tumor cells. Plasma membrane vesicles were isolated from AtT-20 cells and found to take up alpha-aminoisobutyric acid, indicating that they were properly oriented and functionally intact. Corticosterone bound to these vesicle in a time- and temperature-dependent manner. The binding exhibited a glucocorticoid preference since non-glucocorticoids such as progesterone, testosterone or estradiol were unable to inhibit binding. In addition, binding specificity differed from that of the cytoplasmic receptor since the synthetic glucocorticoids were also ineffective competitors. The major inhibitors of binding were corticosterone greater than 11-dehydrocorticosterone greater than 11-ketoprogesterone greater than cortisol. In other complementary studies, AtT-20 cell cytosol was tested to determine whether heterogenous soluble sites exhibiting a preference for the natural vs the synthetic steroid could also be identified. We found that binding sites for both steroid classes were approximately similar in number, specificity and behavior on ion-exchange chromatography. We conclude that, in addition to a classical soluble cytoplasmic glucocorticoid receptor, AtT-20 cells contain plasma membrane glucocorticoid binding sites. The affinity and specificity of these sites for the natural ligand, corticosterone, suggest that they play an important role in the subcellular mechanism of glucocorticoid action.
Immunoselection and tumor evasion constitutes one of the major obstacles in cancer immunotherapy.... more Immunoselection and tumor evasion constitutes one of the major obstacles in cancer immunotherapy. A potential solution to this problem is the development of polyvalent vaccines, and the identification of more tumor-specific antigens is a prerequisite for the development of cancer vaccines. To identify novel tumor-specific antigens, suppression subtractive hybridization (SSH) was performed to isolate genes differentially expressed in human hepatocellular cancer (HCC) tissues. PLAC1 (PLACenta-specific 1) was one of the genes identified highly expressed in HCC tissues but not in paired noncancerous tissues. Further analyses revealed its expression in several other types of cancer tissues as well as tumor cell lines, but not in normal tissues except for placenta. Among HCC samples tested, 32% (22/69) showed PLAC1 mRNA expression while the protein was detected in 23.3% (7/30). A serological survey revealed that 3.8% (4/101) of HCC patients had anti-PLAC1 antibody response, suggesting the immunogenicity of PLAC1 in HCC patients. PLAC1 represents a new class of tumor associated antigen with restricted expression in placenta and cancer tissues, that may serve as a target for cancer vaccination.
Cord sera were obtained from term, Chilean newborns exhibiting various patterns of intrauterine g... more Cord sera were obtained from term, Chilean newborns exhibiting various patterns of intrauterine growth and assayed for IGF-1, IGF-2, IGFBP-1, IGFBP-2, and IGFBP-3 by specific radioimmunoassays (RIA). Serum levels of each peptide were correlated with birth weight (BW), ponderal index (PI), and placental weight (PW). Total IGF-1 levels correlated with BW (r = 0.665, P = 0.0001), PI (r =
Preterm human placentae produce IGF-specific binding proteins. Placental binding proteins are imm... more Preterm human placentae produce IGF-specific binding proteins. Placental binding proteins are immunologically similar to the growth hormone dependent binding protein in human serum (BP-53) and not the growth hormone independent binding protein (BP-28) in human serum and amniotic fluid. Placental production occurs in mesenchymal cells located in the villous core.
Supplementary Data - PDF file 3276K, Supplementary Figure 1. Characterization of MMTV-PPAR{delta}... more Supplementary Data - PDF file 3276K, Supplementary Figure 1. Characterization of MMTV-PPAR{delta} transgenic mice. Supplementary Figure 2. Metabolomic analysis Supplementary Figure 3. Signaling pathways associated with PPRE-containing genes. activated in MMTV-PPAR{delta} mice treated with GW501516 for 11 weeks. Supplementary Table 1. Antibodies for IHC and western blotting. Supplementary Table 2. List of primers for qRT-PCR analysis. Supplementary Table 3. Gene expression preferentially altered in MMTV-PPAR{delta} mice treated with GW501516 and everolimus. Supplementary Table 4. Gene expression preferentially altered in MMTV-PPARd mice treated with GW501516 for 11 weeks. Supplementary Table 5. Gene expression preferentially altered in MMTV-PPARd mice. Supplementary Table 6. Gene expression preferentially altered in wild-type mice treated with GW501516 for 11 weeks.
In previous studies we have found that intact AtT-20 cells contained two glucocorticoid binding s... more In previous studies we have found that intact AtT-20 cells contained two glucocorticoid binding sites with distinctly different affinities and specificity. In this paper, the nature of these sites was investigated by studying glucocorticoid binding to cytosol and to plasma membranes isolated from AtT-20 mouse pituitary tumor cells. Plasma membrane vesicles were isolated from AtT-20 cells and found to take up alpha-aminoisobutyric acid, indicating that they were properly oriented and functionally intact. Corticosterone bound to these vesicle in a time- and temperature-dependent manner. The binding exhibited a glucocorticoid preference since non-glucocorticoids such as progesterone, testosterone or estradiol were unable to inhibit binding. In addition, binding specificity differed from that of the cytoplasmic receptor since the synthetic glucocorticoids were also ineffective competitors. The major inhibitors of binding were corticosterone greater than 11-dehydrocorticosterone greater than 11-ketoprogesterone greater than cortisol. In other complementary studies, AtT-20 cell cytosol was tested to determine whether heterogenous soluble sites exhibiting a preference for the natural vs the synthetic steroid could also be identified. We found that binding sites for both steroid classes were approximately similar in number, specificity and behavior on ion-exchange chromatography. We conclude that, in addition to a classical soluble cytoplasmic glucocorticoid receptor, AtT-20 cells contain plasma membrane glucocorticoid binding sites. The affinity and specificity of these sites for the natural ligand, corticosterone, suggest that they play an important role in the subcellular mechanism of glucocorticoid action.
Immunoselection and tumor evasion constitutes one of the major obstacles in cancer immunotherapy.... more Immunoselection and tumor evasion constitutes one of the major obstacles in cancer immunotherapy. A potential solution to this problem is the development of polyvalent vaccines, and the identification of more tumor-specific antigens is a prerequisite for the development of cancer vaccines. To identify novel tumor-specific antigens, suppression subtractive hybridization (SSH) was performed to isolate genes differentially expressed in human hepatocellular cancer (HCC) tissues. PLAC1 (PLACenta-specific 1) was one of the genes identified highly expressed in HCC tissues but not in paired noncancerous tissues. Further analyses revealed its expression in several other types of cancer tissues as well as tumor cell lines, but not in normal tissues except for placenta. Among HCC samples tested, 32% (22/69) showed PLAC1 mRNA expression while the protein was detected in 23.3% (7/30). A serological survey revealed that 3.8% (4/101) of HCC patients had anti-PLAC1 antibody response, suggesting the immunogenicity of PLAC1 in HCC patients. PLAC1 represents a new class of tumor associated antigen with restricted expression in placenta and cancer tissues, that may serve as a target for cancer vaccination.
Cord sera were obtained from term, Chilean newborns exhibiting various patterns of intrauterine g... more Cord sera were obtained from term, Chilean newborns exhibiting various patterns of intrauterine growth and assayed for IGF-1, IGF-2, IGFBP-1, IGFBP-2, and IGFBP-3 by specific radioimmunoassays (RIA). Serum levels of each peptide were correlated with birth weight (BW), ponderal index (PI), and placental weight (PW). Total IGF-1 levels correlated with BW (r = 0.665, P = 0.0001), PI (r =
Preterm human placentae produce IGF-specific binding proteins. Placental binding proteins are imm... more Preterm human placentae produce IGF-specific binding proteins. Placental binding proteins are immunologically similar to the growth hormone dependent binding protein in human serum (BP-53) and not the growth hormone independent binding protein (BP-28) in human serum and amniotic fluid. Placental production occurs in mesenchymal cells located in the villous core.
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