Journal: PLoS ONE

Article Title: GSK3β Inhibition Promotes Synaptogenesis in Drosophila and Mammalian Neurons

doi: 10.1371/journal.pone.0118475

Figure Lengend Snippet: A. Immunoblot of p-β-Catenin from 12 DIV hippocampal neurons treated for 48 hours with PTD4-PI3KAc or PTD4 (21 μM). B. Immunoblot of p-β-Catenin after AR-A014418 treatment, alone or in combination with PTD4-PI3KAc (21 μM). Phosphorylation levels were normalized to the total actin. C. Plot bar representing the mean average of p-β-Catenin after the described treatment. All percentages are referred to control conditions as 100%, always included in each Western-blot membrane. Only the treatment with 2 and 10 μM of AR-A014418 decreased p-β-Catenin levels. The co-treatment with the PI3K activator did not induce a further reduction. D. Immunoblot of p-β-Catenin after SB 415286 or in combination with PTD4-PI3KAc (21 μM) treatments. E. Plot bar of the percentage of p-β-Catenin after treatment with SB 415286. Exposure of cells to SB 415286 or in combination with PI3K activator led to a statistically significant decrease in p-β-Catenin (n = 15 from 15 different cultures). Percentage of p-β-Catenin was normalized to the total Actin levels. Significance refers to the statistical analysis of the experiments versus the control (100%). Student’s two-tailed t-test, *p<0.05, **p<0.005.

Article Snippet: GSK3 inhibitors: SB-415286 and AR-A014418 [ – ] were from Tocris, UK (ref. 1617) and from Sigma, USA (ref. A3230), respectively.

Techniques: Western Blot, Phospho-proteomics, Control, Membrane, Two Tailed Test

Journal: PLoS ONE

Article Title: GSK3β Inhibition Promotes Synaptogenesis in Drosophila and Mammalian Neurons

doi: 10.1371/journal.pone.0118475

Figure Lengend Snippet: A. Example of a 21 DIV hippocampal neuron in culture. Scale bar = 10 μm. Bottom pictures: detailed magnification of the area depicted by the white box. Synapses (a) were localized following a double criterion: puncta positive for Synapsin (b, red) and for Bassoon (c, green). Scale bar 3 μm. B. Plot graph of the average changes in synaptic density at 21 DIV after AR-A014418 treatment. PTD4-PI3KAc and PTD4 were used at 21 μM. C. Plot graph of the results of synaptic density after the GSK3 inhibition by SB 415286 treatment. D. Quantification of synaptic density at 12 DIV after treatments with PTD4-PI3KAc, AR-A014418, SB 415286 alone or in combination with PTD4-PI3KAc. Percentage of change refers to PTD4 control treatment. Cultures were treated for 48 hours before fixation (n = 10–12 from 3 different cultures). Significance refers to the statistical analysis of each experimental condition versus PTD4 treated cultures (100%) Student’s two-tailed t-test, *p<0.05, **p<0.005, ***p<0.0001.

Article Snippet: GSK3 inhibitors: SB-415286 and AR-A014418 [ – ] were from Tocris, UK (ref. 1617) and from Sigma, USA (ref. A3230), respectively.

Techniques: Inhibition, Control, Two Tailed Test

Journal: PLoS ONE

Article Title: GSK3β Inhibition Promotes Synaptogenesis in Drosophila and Mammalian Neurons

doi: 10.1371/journal.pone.0118475

Figure Lengend Snippet: A. Representative immunoblot showing Synapsin levels after GSK3 inhibition by SB 415286 treatment in 21 DIV neuronal hippocampal cultures. B. Plot graph summary of the results of 21 DIV cultures. The bars are the average levels of Synapsin versus control conditions after the indicated treatments, alone or in combination with PTD4-PI3KAc (21 μM). C. Representative immunoblot for Synapsin after GSK3 inhibition in 12 DIV cultures D. Plot graph of Synapsin after SB 415286 treatment, alone or in combination with PTD4-PI3KAc (21 μM). E. Representative immunoblot of Synapsin after GSK3 inhibition by AR-A014418 in 12 DIV cultures. F. Plot bars of the average levels of Synapsin after AR-A014418 treatment, alone or in combination with PTD4-PI3KAc (21 μM). Percentage of Synapsin was normalized to the total Actin levels. n = 10–15 from 6 different cultures. Significance refers to the statistical analysis of the experiments versus the control (100%). Student’s two-tailed t-test, *p<0.05, **p<0.005, ***p<0.0001.

Article Snippet: GSK3 inhibitors: SB-415286 and AR-A014418 [ – ] were from Tocris, UK (ref. 1617) and from Sigma, USA (ref. A3230), respectively.

Techniques: Western Blot, Inhibition, Control, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Representative images showing type I fibers (I, green) and type II fibers (II, dark) in rat soleus muscle. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418). Scale bar, 100 μm.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on GSK-3β Ser9 phosphorylation ( a ) and GS-1 Ser 641 phosphorylation ( b ) in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C, $: p < 0.05 vs. 7HS.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on β-catenin content in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C, $: p < 0.05 vs. 7HS.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on the amount of 18S rRNA ( a ) and 28S rRNA ( b ) in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C, $: p < 0.05 vs. 7HS.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on the expression of 45S pre-rRNA ( a ) and c-Myc ( b ) in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C, $: p < 0.05 vs. 7HS.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on rpS6 abundance ( a ) and phosphorylation status of rpS6 ( b ) and 4E-BP1 ( c ) in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: The Role of Glycogen Synthase Kinase-3 in the Regulation of Ribosome Biogenesis in Rat Soleus Muscle under Disuse Conditions

doi: 10.3390/ijms23052751

Figure Lengend Snippet: Effect of GSK-3 inhibition during 7-day unloading on the rate of protein synthesis in rat soleus muscle. Data are presented as median, interquartile range, minimum and maximum values, expressed as % of control. C, vivarium control group, C + AR, vivarium control rats + daily injections of GSK-3 inhibitor (AR-A014418) for 7 days, 7HS, hindlimb suspension for 7 days, 7HS + AR, 7-day hindlimb suspension + daily injections of GSK-3 inhibitor (AR-A014418), *: p < 0.05 vs. C, &: p < 0.05 vs. 7HS.

Article Snippet: AR-A014418 (#A3184; APExBIO, Houston, TX, USA) was administered once a day (each injection contained 4 mg/kg in saline + 1% DMSO).

Techniques: Inhibition