Journal: Cell Death & Disease

Article Title: PTEN regulates AMPA receptor-mediated cell viability in iPS-derived motor neurons

doi: 10.1038/cddis.2014.55

Figure Lengend Snippet: PTEN knockdown ( b and f ) increases phosphorylation of AKT ( c and g ) and Bad ( d and h ) in primary cultured motor neurons and iPS-derived motor neurons. Primary ( a ) or iPS-derived ( e ) motor neurons were immunostained with anti-tubulin (green), Hoechst (nuclear staining in blue) and PTEN (red). Scale bar, 20 μ m. Western blotting on homogenates from primary cultured ( b–d ) and iPS-derived motor neurons. PTEN is normalized by GAPDH ( b and f ) and pAKT is normalized by AKT ( c and g ). pBAD is normalized by BAD ( d and h ). Results represent the mean+S.E.M. from four independent experiments. ** P <0.01, tested by one-way ANOVA

Article Snippet: For iPS-derived MNs, PTEN shRNA (h) Lentiviral vector containing three target-specific constructs that encode 19–25 nt (plus hairpin) shRNA designed to target human PTEN were purchased from Santa Cruz Biotechnology (sc-29459 and control shRNA Lentiviral Particles: sc-108080).

Techniques: Knockdown, Phospho-proteomics, Cell Culture, Derivative Assay, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: PTEN regulates AMPA receptor-mediated cell viability in iPS-derived motor neurons

doi: 10.1038/cddis.2014.55

Figure Lengend Snippet: PTEN knockdown decreases AMPA receptor expression in primary cultured ( a – f ) and iPS-derived ( g – l ) motor neurons. Motor neurons were immunostained with anti-tubulin (green), Hoechst (nuclear staining in blue) and AMPA receptor (GluR1, red). Scale bar, 20 μ m. ( b – f and h – l ) Western blotting. PTEN knockdown decreases GluR1 ( c and i ) and GluR2 ( d and j ) but not GluR4 expression in both primary cultured and iPS-derived MNs. GluR3 decreases significantly in iPS-derived MNs ( k ) but not in primary cultured MNs ( e ). Results represent the mean+S.E.M. from four independent experiments. ** P <0.01, tested by one-way ANOVA

Article Snippet: For iPS-derived MNs, PTEN shRNA (h) Lentiviral vector containing three target-specific constructs that encode 19–25 nt (plus hairpin) shRNA designed to target human PTEN were purchased from Santa Cruz Biotechnology (sc-29459 and control shRNA Lentiviral Particles: sc-108080).

Techniques: Knockdown, Expressing, Cell Culture, Derivative Assay, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: PTEN regulates AMPA receptor-mediated cell viability in iPS-derived motor neurons

doi: 10.1038/cddis.2014.55

Figure Lengend Snippet: PTEN knockdown decreases AMPA-induced whole-cell currents in primary cultured ( a – d ) and iPS-derived ( e – h ) motor neurons. Primary cultured ( a ) and iPS-derived ( e ) motor neurons were immunostained with anti-tubulin (green), Hoechst (nuclear staining in blue) and ChAT (red). Scale bar, 20 μ m. ( b and f ) Representative current traces elicited by AMPA. ( c and g ) Dose response of AMPA-induced whole-cell currents were recorded in 20 mM extracellular Na+ at −60 mV, in response to AMPA concentrations ranging from 5 μ M to 5 mM. Each point represents the mean±S.E.M. from three cells (* P <0.05, ** P <0.01, tested by one-way ANOVA). EC 50 values estimated from fits to pooled data were 106 μ M for primary cultured motor neurons and 113 μ M for iPS-derived neurons. ( d and h ) Average peak AMPA-induced whole-cell currents were recorded in Na+-free extracellular solution containing 50 mM Ca 2+ at −60 mV in primary cultured and iPS-derived motor neurons, evoked by AMPA 100 μ M ( n =8). Columns represent the peak amplitudes of agonist-induced whole-cell currents. Results represent the mean+S.E.M. from four independent experiments, with 8 motor neurons of each (* P <0.05, ** P <0.01, tested by one-way ANOVA)

Article Snippet: For iPS-derived MNs, PTEN shRNA (h) Lentiviral vector containing three target-specific constructs that encode 19–25 nt (plus hairpin) shRNA designed to target human PTEN were purchased from Santa Cruz Biotechnology (sc-29459 and control shRNA Lentiviral Particles: sc-108080).

Techniques: Knockdown, Cell Culture, Derivative Assay, Staining

Journal: Cell Death & Disease

Article Title: PTEN regulates AMPA receptor-mediated cell viability in iPS-derived motor neurons

doi: 10.1038/cddis.2014.55

Figure Lengend Snippet: PTEN knockdown decreases AMPA-induced cell death in primary cultured and iPS-derived motor neurons. Primary cultured ( a ) and iPS-derived ( c ) motor neurons were immunostained with anti-tubulin (green), Hoechst (nuclear staining in blue) and TUNEL staining (red). Scale bar, 20 μ m. ( b and d ) Results represent the mean+S.E.M. from four independent experiments. ** P <0.01, tested by one-way ANOVA

Article Snippet: For iPS-derived MNs, PTEN shRNA (h) Lentiviral vector containing three target-specific constructs that encode 19–25 nt (plus hairpin) shRNA designed to target human PTEN were purchased from Santa Cruz Biotechnology (sc-29459 and control shRNA Lentiviral Particles: sc-108080).

Techniques: Knockdown, Cell Culture, Derivative Assay, Staining, TUNEL Assay