Journal: International Journal of Molecular Sciences

Article Title: miR-132-3p Modulates DUSP9-Dependent p38/JNK Signaling Pathways to Enhance Inflammation in the Amnion Leading to Labor

doi: 10.3390/ijms23031864

Figure Lengend Snippet: The role of p38 and JNK in DUSP9 siRNA-induced expression of proinflammatory cytokines and COX2 as well as PGE2 in WISH cells. WISH cells were transfected with si- DUSP9 or NC for 24 h, followed by 24 h of treatment with DMSO, p38 inhibitor SB203580 (10 μM), or JNK inhibitor SP600125 (20 μM). RT-qPCR analysis of the expression of IL-1β , TNF-α , COX2 ( A , G ), IL-6 , and IL-8 ( B , H ). Magnetic Luminex Assays of the secretion of IL-1β, TNF-α ( C , I ), IL-6, and IL-8 ( D , J ). Western blot analysis of COX2 expression ( E , K ). ELISA analysis of PGE2 level ( F , L ). Data were shown as mean ± SEM, n = 3. * p < 0.05, ** p < 0.01 vs. control (0), # p < 0.05, ## p < 0.01 vs. si- DUSP9 .

Article Snippet: To verify the involvement of p38 and JNK signaling pathways in the induction of IL-1β, IL-6, IL-8, TNF-α, and COX2 as well as PGE2 secretion by DUSP9 siRNA, the cells were transfected with NC or si- DUSP9 for 24 h, followed by 24 h of treatment with dimethyl sulfoxide (DMSO), p38 inhibitor SB203580 (10 μM, Selleck, Houston, TX, USA), or JNK inhibitor SP600125 (20 μM, Selleck, Houston, TX, USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Luminex, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: The Journal of Neuroscience

Article Title: Abl Tyrosine Kinase Promotes Dendrogenesis by Inducing Actin Cytoskeletal Rearrangements in Cooperation with Rho Family Small GTPases in Hippocampal Neurons

doi: 10.1523/JNEUROSCI.1264-04.2004

Figure Lengend Snippet: Immunofluorescent images of hippocampal neurons at 5 DIV treated with 0.03% DMSO (left) or 3 μm STI571 (right) for 48 hr and double-immunolabeled with anti-MAP2 and anti-CD71 (transferrin receptor) or anti-NR2A and anti-Tau. Arrows point to the dendrites; arrowhead points to the axon. Note the colocalization of MAP2 with CD71 in DMSO- and STI571-treated cells. Also note that, whereas Tau was localized to the axon, NR2A was localized more prominently in dendrites (arrow), with weaker staining in the axon (DMSO; compare Tau and NR2A staining). The segregation of Tau and NR2A was not clear in neurons treated with STI571. Scale bar, 15 μm.

Article Snippet: Alternatively, neurons were subject to 100 n m jasplakinolide (Molecular Probes, Eugene, OR) for 5 hr, 5 μ m vincristine (Sigma, St. Louis, MO) for 5 hr, 5 μ m latrunculin A (Biomol Research Laboratory, Plymouth Meeting, PA) treatment for 24 hr, or 0.03% DMSO (Sigma) treatment for 24 hr before they were fixed for immunofluorescent light microscopy.

Techniques: Immunolabeling, Staining

Journal: The Journal of Neuroscience

Article Title: Abl Tyrosine Kinase Promotes Dendrogenesis by Inducing Actin Cytoskeletal Rearrangements in Cooperation with Rho Family Small GTPases in Hippocampal Neurons

doi: 10.1523/JNEUROSCI.1264-04.2004

Figure Lengend Snippet: Abl family tyrosine kinase activity influences primary dendrite formation and dendritic branching. A, Phase-contrast and double-immunofluorescent light microscopy of single neurons treated with DMSO or STI571 or transfected with CA-Abl. Neurons were treated for 48 hr from 5 to 7 DIV and immunostained with rabbit polyclonal anti-Abl and mouse monoclonal anti-MAP2. Note the simplification of dendritic profile in STI571-treated neurons in comparison to an exuberant outgrowth of MAP2-positive, highly branched processes in neurons transfected with CA-Abl. Scale bar, 15 μm. B, Quantification of primary and secondary dendrite outgrowth in neurons incubated with DMSO or STI571 or transfected with CA-Abl. C, Quantification of primary and secondary dendrite length in neurons incubated with DMSO or STI571 or transfected with CA-Abl. Error bars indicate SEM; *p < 0.05.

Article Snippet: Alternatively, neurons were subject to 100 n m jasplakinolide (Molecular Probes, Eugene, OR) for 5 hr, 5 μ m vincristine (Sigma, St. Louis, MO) for 5 hr, 5 μ m latrunculin A (Biomol Research Laboratory, Plymouth Meeting, PA) treatment for 24 hr, or 0.03% DMSO (Sigma) treatment for 24 hr before they were fixed for immunofluorescent light microscopy.

Techniques: Activity Assay, Light Microscopy, Transfection, Incubation

Journal: The Journal of Neuroscience

Article Title: Abl Tyrosine Kinase Promotes Dendrogenesis by Inducing Actin Cytoskeletal Rearrangements in Cooperation with Rho Family Small GTPases in Hippocampal Neurons

doi: 10.1523/JNEUROSCI.1264-04.2004

Figure Lengend Snippet: ROCK inhibition suppresses STI571 effect on dendrogenesis. A, Immunofluorescent light microscopy of anti-MAP2 and rhodamine phalloidin staining of 5 DIV hippocampal neurons treated with DMSO (STI571/- and Y-27632/-) or 3 μm STI571 for 48 hr or treated with 3 μm STI571 for 24 hr, followed by the addition of Y-27632 for an additional 18 hr. Cells were fixed on 7 DIV. Arrowheads point to focal actin filaments. Scale bar, 15 μm. B, Comparison of the percentage of change of dendrite numbers among neurons treated with DMSO, STI571, Y-27632, and STI571/Y-27632. Note that the reduction of dendrite numbers by STI571 treatment is reversed by coincubation of neurons with Y-27632. C, Comparison of the percentage of change of dendrite length among neurons treated with DMSO, STI571, Y-27632, and STI571/Y-27632. Note that the decrease in dendrite length by STI571 treatment is reversed by coincubation of neurons with Y-27632. The effect of Y-27632 is more dramatic on the primary dendrites than on the secondary dendrites in neurons 7 DIV. Error bars indicate SEM; *p < 0.05.

Article Snippet: Alternatively, neurons were subject to 100 n m jasplakinolide (Molecular Probes, Eugene, OR) for 5 hr, 5 μ m vincristine (Sigma, St. Louis, MO) for 5 hr, 5 μ m latrunculin A (Biomol Research Laboratory, Plymouth Meeting, PA) treatment for 24 hr, or 0.03% DMSO (Sigma) treatment for 24 hr before they were fixed for immunofluorescent light microscopy.

Techniques: Inhibition, Light Microscopy, Staining

Journal: The Journal of Neuroscience

Article Title: Abl Tyrosine Kinase Promotes Dendrogenesis by Inducing Actin Cytoskeletal Rearrangements in Cooperation with Rho Family Small GTPases in Hippocampal Neurons

doi: 10.1523/JNEUROSCI.1264-04.2004

Figure Lengend Snippet: The CA-Abl effect on dendrogenesis is dependent on the reorganization of actin cytoskeleton but is independent of the stability of microtubules. A, Immunofluorescent light microscopy of single neurons by anti-Abl antibody (top) and rhodamine phalloidin (bottom). The 5 DIV hippocampal neurons transfected with CA-Abl were treated either with 5 μm latrunculin A the next day for 24 hr to depolymerize actin filament or with 100 nm jasplakinolide on 7 DIV for 5 hr to stabilize actin filaments. Neurons were fixed on 7 DIV. Note that CA-Abl-induced process outgrowth is suppressed by jasplakinolide. In jasplakinolide-treated neurons, F-actin staining is weak because of competitive binding of jasplakinolide and phalloidin to F-actin. Note that F-actin staining in latrunculin A-treated neurons was enhanced to quantify the processes. Scale bar, 15 μm. B, Quantification of primary and secondary dendrite outgrowth in neurons incubated with DMSO, latrunculin A, and jasplakinolide as well as transfected with CA-Abl in the presence or absence of latrunculin A and jasplakinolide. C, Quantification of the relative length of primary and secondary dendrites in neurons incubated with DMSO, latrunculin A, and jasplakinolide as well as transfected with CA-Abl in the presence and absence of latrunculin A and jasplakinolide. Dendrite number and length were determined within 120 μm2 of the single neuronal cell bodies. Data represent the means ± SEM of three independent experiments; *p < 0.05.

Article Snippet: Alternatively, neurons were subject to 100 n m jasplakinolide (Molecular Probes, Eugene, OR) for 5 hr, 5 μ m vincristine (Sigma, St. Louis, MO) for 5 hr, 5 μ m latrunculin A (Biomol Research Laboratory, Plymouth Meeting, PA) treatment for 24 hr, or 0.03% DMSO (Sigma) treatment for 24 hr before they were fixed for immunofluorescent light microscopy.

Techniques: Light Microscopy, Transfection, Staining, Binding Assay, Incubation