Immunoblotting

Publication Title: 
Molecular Cell

Cellular processes function through multistep pathways that are reliant on the controlled association and disassociation of sequential protein complexes. While dynamic action is critical to propagate and terminate work, the mechanisms used to disassemble biological structures are not fully understood. Here we show that the p23 molecular chaperone initiates disassembly of protein-DNA complexes and that the GCN5 acetyltransferase prolongs the dissociated state through lysine acetylation.

Author(s): 
Zelin, Elena
Zhang, Yang
Toogun, Oyetunji A.
Zhong, Sheng
Freeman, Brian C.
Publication Title: 
Archives of General Psychiatry

BACKGROUND: Schizophrenia is frequently accompanied by hypometabolism and altered gene expression in the prefrontal cortex. Cellular metabolism regulates chromatin structure, including covalent histone modifications, which are epigenetic regulators of gene expression. OBJECTIVE: To test the hypothesis that down-regulated metabolic gene expression is associated with histone modification changes in the prefrontal cortex of subjects with schizophrenia.

Author(s): 
Akbarian, Schahram
Ruehl, Martin G.
Bliven, Erin
Luiz, Lori A.
Peranelli, Amy C.
Baker, Stephen P.
Roberts, Rosalinda C.
Bunney, William E.
Conley, Robert C.
Jones, Edward G.
Tamminga, Carol A.
Guo, Yin
Publication Title: 
Molecular Cancer Therapeutics

The present study was undertaken to gain insights into the molecular mechanism of cell death (apoptosis) by guggulsterone, a constituent of Ayurvedic medicinal plant Commiphora mukul, using PC-3 human prostate cancer cells as a model. The viability of PC-3 cells, but not a normal prostate epithelial cell line (PrEC), was reduced significantly on treatment with guggulsterone in a concentration-dependent manner.

Author(s): 
Singh, Shivendra V.
Zeng, Yan
Xiao, Dong
Vogel, Victor G.
Nelson, Joel B.
Dhir, Rajiv
Tripathi, Yamini B.
Publication Title: 
Molecular Cancer Therapeutics

The present study was undertaken to gain insights into the molecular mechanism of cell death (apoptosis) by guggulsterone, a constituent of Ayurvedic medicinal plant Commiphora mukul, using PC-3 human prostate cancer cells as a model. The viability of PC-3 cells, but not a normal prostate epithelial cell line (PrEC), was reduced significantly on treatment with guggulsterone in a concentration-dependent manner.

Author(s): 
Singh, Shivendra V.
Zeng, Yan
Xiao, Dong
Vogel, Victor G.
Nelson, Joel B.
Dhir, Rajiv
Tripathi, Yamini B.
Publication Title: 
Mutation Research

Since trivalent chromium (Cr(3+)) enhances glucose metabolism, interest in the use of Cr(3+)as a therapy for type 2 diabetes has grown in the mainstream medical community. Moreover, accumulating evidence suggests that Cr(3+) may also benefit cardiovascular disease (CVD) and atypical depression. We have found that cholesterol, a lipid implicated in both CVD and neurodegenerative disorders, also influences cellular glucose uptake. A recent study in our laboratory shows that exposure of 3T3-L1 adipocytes to chromium picolinate (CrPic, 10 nM) induces a loss of plasma membrane cholesterol.

Author(s): 
Pattar, Guruprasad R.
Tackett, Lixuan
Liu, Ping
Elmendorf, Jeffrey S.
Publication Title: 
Brain Research

MAP kinase is associated with delta-opioid receptor (DOR) signaling and plays a role in cell survival/death. Since anisomycin may alter MAP kinase activity and affect neuronal survival, we investigated whether anisomycin alters neuronal response to hypoxic stress and DOR inhibition. The experiments were performed in cultured cortical neurons. MAP kinase activities were determined by immunoblotting and neuronal viability was assessed by LDH leakage and live/dead morphological study.

Author(s): 
Hong, Soon-Sun
Qian, Hong
Zhao, Peng
Bazzy-Asaad, Alia
Xia, Ying
Publication Title: 
Molecular Endocrinology (Baltimore, Md.)

Previously, we found that a loss of plasma membrane (PM) phosphatidylinositol 4,5-bisphosphate (PIP2)-regulated filamentous actin (F-actin) structure contributes to insulin-induced insulin resistance. Interestingly, we also demonstrated that chromium picolinate (CrPic), a dietary supplement thought to improve glycemic status in insulin-resistant individuals, augments insulin-regulated glucose transport in insulin-sensitive 3T3-L1 adipocytes by lowering PM cholesterol.

Author(s): 
Horvath, Emily M.
Tackett, Lixuan
McCarthy, Alicia M.
Raman, Priya
Brozinick, Joseph T.
Elmendorf, Jeffrey S.
Publication Title: 
Toxicology and Applied Pharmacology

CD40 is a costimulatory molecule linking innate and adaptive immune responses to bacterial stimuli, as well as a critical regulator of functions of other costimulatory molecules. The mechanisms regulating lipopolysaccharide (LPS)-induced CD40 expression have not been adequately characterized in human monocytic cells. In this study we used a human monocytic cell line, THP-1, to investigate the possible mechanisms of CD40 expression following LPS exposure. Exposure to LPS resulted in a dose- and time-dependent increase in CD40 expression.

Author(s): 
Wu, Weidong
Alexis, Neil E.
Chen, Xian
Bromberg, Philip A.
Peden, David B.
Publication Title: 
Biochimica Et Biophysica Acta

Ceramide is an important bioactive lipid, intimately involved in many cellular functions, including the regulation of cell death, and in cancer and chemotherapy. Ceramide is synthesized de novo from sphinganine and acyl CoA via a family of 6 ceramide synthase enzymes, each having a unique preference for different fatty acyl CoA substrates and a unique tissue distribution. However, little is known regarding the regulation of these important enzymes.

Author(s): 
Sridevi, Priya
Alexander, Hannah
Laviad, Elad L.
Pewzner-Jung, Yael
Hannink, Mark
Futerman, Anthony H.
Alexander, Stephen
Publication Title: 
Cancer Research

The CXC receptor-1 (CXCR1) is a coreceptor for interleukin-8 (IL-8) and is expressed on both normal and tumor cells. The function of CXCR1 in prostate cancer was investigated by silencing its expression, using RNA interference. We established stable cell colonies of PC-3 cells, depleted of CXCR1, using lentiviral plasmids (pLK0.1puro) generating small hairpin RNA (shRNA) against CXCR1 mRNA. Stable shRNA transfectants (PLK1-PLK5) that express significantly reduced CXCR1 mRNA (>or=90% down) and protein (>or=43% down) or vector-only transfectants (PC-3V) were characterized.

Author(s): 
Shamaladevi, Nagarajarao
Lyn, Dominic A.
Escudero, Diogo O.
Lokeshwar, Bal L.

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