Journal: ACS chemical neuroscience

Article Title: Chronic Social Isolation Stress during Peri-Adolescence Alters Presynaptic Dopamine Terminal Dynamics via Augmentation in Accumbal Dopamine Availability

doi: 10.1021/acschemneuro.8b00360

Figure Lengend Snippet: Assessment of several terminal protein expression levels. Relative protein expression levels of (A) VMAT2 (n = 7 in both groups), (B) Synaptigyrin-3 (aGH: n = 6; aSI: n = 5), (C) Syntaxin-1 (aGH: n = 7; aSI: n = 6), and (D) Munc13-3 (aGH: n = 8; aSI: n = 8) were measured. None of these proteins had different expression levels in aGH and aSI rats. (insets) The representative Western blot images, with the respective protein and actin for comparison. Group housed, aGH, blue; Socially isolated, aSI, red.

Article Snippet: Subsequently, blots were incubated with agitation for 2 h at room temperature in TBS-T/5% bovine serum albumin (05470; Sigma-Aldrich) solution containing the following primary antibody concentrations: VAMT2 (1:2000; AB1598P; Millipore Sigma); Synaptogyrin-3 (1:1000; ab106460; abcam); Syntaxin-1 (1:1000; ANR-002; Alomone laboratories).

Techniques: Expressing, Western Blot, Isolation

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Roles of Id1/HIF-1 and CDK5/HIF-1 in cell cycle reentry induced by amyloid-beta peptide in post-mitotic cortical neuron.

doi: 10.1016/j.bbamcr.2019.118628

Figure Lengend Snippet: Fig. 3. Aβ25-35 induces p25 production from cleavage of p35 without affecting CDK5 expression in cultured cortical neurons. Primary cortical cultures were treated with 10 μM Aβ25-35 for indicated times before detection of p35/p25 (A) or CDK5 (E) by Western blotting. The β-actin in (A) and α-tubulin in (E) served as the respective internal control for equal loading of proteins in each lane. Quantitative analyses of the signal intensities on the blots for p25, p35, p25/p35 ratio, and CDK5 are shown respectively in (B), (C), (D), and (F). Mean ± S.E.M. from N = 3 in all the statistical analyses. * denotes P < 0.05 compared with the corresponding control cultures without Aβ25-35 treatment in (B)–(D). The “ns” in (F) denotes “not significant”.

Article Snippet: The mouse antibody against β-actin (1:4000; Cat. No. MAB1501; Merck Millipore), mouse antibody against α-tubulin (1:10000; Cat. No. T9026; Sigma-Aldrich), rabbit antibody against cyclin D1 (1:1000; Cat. No. ab134175; Abcam), rabbit antibody against PCNA (1:500; Cat. No. ab92552; Abcam), rabbit antibody against HIF-1α (1:500; Cat. No. NB100-134; Novus Biologicals, Littleton, CO, USA), rabbit antibody against CDK5 (1:500; Cat. No. ab40773; Abcam), and rabbit antibody against p35/p25 (1:500; Cat. No. 2680; Cell Signaling Technology, Beverly, MA, USA) were all diluted in blocking buffer (5% nonfat dry milk in TBST buffer containing 0.05% Tween 20, 137 mM NaCl, and 20 mM Tris–HCl, pH 7.5).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Roles of Id1/HIF-1 and CDK5/HIF-1 in cell cycle reentry induced by amyloid-beta peptide in post-mitotic cortical neuron.

doi: 10.1016/j.bbamcr.2019.118628

Figure Lengend Snippet: Fig. 5. Suppression of Id1 enhances p25 production, whereas exogenous Id1-Tag suppresses p25 production in rat cortical cultures. (A–F) Primary cortical cultures were transfected with sc siRNA or Id1 siRNA for 72 h followed by exposure to 10 μM Aβ25-35 for additional 4 h before detection of Id1 (A) or p35/p25 (C) by Western blotting. Quantitative analyses of the signal intensities on the blots for Id1, p25, p25/p35 ratio, and p35 are respectively shown in (B), (D), (E), and (F). Mean ± S.E.M. from N = 3 in all the statistical analyses. * and # denote P < 0.05. The “ns” denotes “not significant”. (G–L) Primary cortical cultures were treated with or without 4 μg/ml Id1-Tag for 2 h or 4 h before detection of p35/p25 (G) or CDK5 (K). Quantitative analyses of the signal intensities on the blots for p25, p25/ p35 ratio, p35, and CDK5 are shown respectively in (H), (I), (J), and (L). Mean ± S.E.M. from N = 4 in 2 h and N = 5 in 4 h. * denotes P < 0.05 as compared to the corresponding control cultures. The “ns” denotes “not significant”.

Article Snippet: The mouse antibody against β-actin (1:4000; Cat. No. MAB1501; Merck Millipore), mouse antibody against α-tubulin (1:10000; Cat. No. T9026; Sigma-Aldrich), rabbit antibody against cyclin D1 (1:1000; Cat. No. ab134175; Abcam), rabbit antibody against PCNA (1:500; Cat. No. ab92552; Abcam), rabbit antibody against HIF-1α (1:500; Cat. No. NB100-134; Novus Biologicals, Littleton, CO, USA), rabbit antibody against CDK5 (1:500; Cat. No. ab40773; Abcam), and rabbit antibody against p35/p25 (1:500; Cat. No. 2680; Cell Signaling Technology, Beverly, MA, USA) were all diluted in blocking buffer (5% nonfat dry milk in TBST buffer containing 0.05% Tween 20, 137 mM NaCl, and 20 mM Tris–HCl, pH 7.5).

Techniques: Transfection, Western Blot, Control

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Roles of Id1/HIF-1 and CDK5/HIF-1 in cell cycle reentry induced by amyloid-beta peptide in post-mitotic cortical neuron.

doi: 10.1016/j.bbamcr.2019.118628

Figure Lengend Snippet: Fig. 7. A diagram showing the proposed signal transduction pathway. We propose possible molecular mechanisms un- derlying Aβ25-35-dependent neuronal cell cycle reentry through Id1- and CDK5/p25-dependent HIF-1α induction in the fully differentiated post-mitotic cortical neurons. This mechanism is further complicated by the intriguing antag- onism between Id1 and p25 because they reciprocally sup- press expression of each other. Possible involvements of unknown mediators, as indicated by a red question mark, linking Aβ and HIF-1 is also shown. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The mouse antibody against β-actin (1:4000; Cat. No. MAB1501; Merck Millipore), mouse antibody against α-tubulin (1:10000; Cat. No. T9026; Sigma-Aldrich), rabbit antibody against cyclin D1 (1:1000; Cat. No. ab134175; Abcam), rabbit antibody against PCNA (1:500; Cat. No. ab92552; Abcam), rabbit antibody against HIF-1α (1:500; Cat. No. NB100-134; Novus Biologicals, Littleton, CO, USA), rabbit antibody against CDK5 (1:500; Cat. No. ab40773; Abcam), and rabbit antibody against p35/p25 (1:500; Cat. No. 2680; Cell Signaling Technology, Beverly, MA, USA) were all diluted in blocking buffer (5% nonfat dry milk in TBST buffer containing 0.05% Tween 20, 137 mM NaCl, and 20 mM Tris–HCl, pH 7.5).

Techniques: Transduction, Expressing