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Fig. 2 | Molecular Autism

Fig. 2

From: Cytoplasm-predominant Pten associates with increased region-specific brain tyrosine hydroxylase and dopamine D2 receptors in mouse model with autistic traits

Fig. 2

PTEN suppresses TH expression via inhibition of the PI3K/CREB pathway. a FLAG-tagged wild-type PTEN was expressed (Tet−, i.e., Tet-Off) or not expressed (Tet+) in PC12 cells, whose lysates were interrogated by Western blot for the labeled proteins/phosphoproteins (left panel). GAPDH is loading control. Note that FLAG-PTEN expression associates with decreased P-AKT (right lane vs left lane), decreased P-CREB, and decreased TH. Normalized quantitation of TH and P-CREB in the presence of PTEN overexpression (Tet−) or no PTEN overexpression (Tet+) (right panel). Note that Tet+ PC12 cells have endogenous rat wild-type Pten. b Western blots for named proteins/P-proteins after inhibition of PI3K by LY294002 (left panel) or serum starvation (middle panel) and after upregulation of PI3K by brief NGF exposure (right panel). Note that PI3K inhibition (manifested by increased P-AKT) is associated with decreased phospho-CREB and TH, whereas NGF stimulation of PI3K (manifested by decreased P-AKT) is associated with increased P-CREB and TH expression. c Western blot for TH, P-CREB, CREB, and GAPDH loading control after inhibition of CREB phosphorylation by expression of a CREB dominant-negative mutant (K-CREB) and a phosphorylation mutant (CREB S133A) compared to wild-type CREB (CREB WT). Note decreased TH expression associated with both (inactive) CREB mutants. d Identification of a CREB consensus binding site at the 5′UTR of the TH gene. e Elevated CREB phosphorylation in the cerebrum from Pten m3m4/m3m4 mutant mice

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