instat prism 7 software  (GraphPad Software Inc)

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced oxidative stress in BV2 microglial cells. The BV2 and BV2-G microglial cells were seeded (3×106) in a 96-well plate. Then the cells were pre-incubated with DCFH-DA dye for 45 min and the cells were treated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were used to measure ROS intensity on a fluorescence microscope. Representative images show the toxic effect of MPP+ induced ROS intensity (green fluorescence; A). Bar graphs show the green fluorescence intensity significantly reduced in GMF deficient BV2-G cells as compared with BV2 microglial cells (B). Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. The p value less than <0.05 was considered as statistically significant in all the experiments; Values are given as mean ± SEM of three experiments in each group. p<0.001 and p<0.01 untreated controls vs MPP+-treated cells; p<0.05 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. Scale bar 100 µm. AU arbitrary unit.

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Incubation, Fluorescence, Microscopy, Software

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced apoptosis in BV2 microglial cells. The BV2 and BV2-G cells were seeded (3×106) in a 96-well plate. The cells were treated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were stained with EtBr/AO solution (1:1 v/v) at a final concentration of 100 µg/ml for 5 min and finally imaged under fluorescence microscope. Images represents the toxic effect of MPP+ -induced apoptotic cell death (dark orange red; Fig 2A). Bar graphs show that apoptotic changes were significantly reduced in BV2-G microglial cells as compared with BV2 cells (B). Values are given as mean ± SEM of three experiments in each group. Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. p<0.001 untreated controls vs MPP+-treated cells; p<0.01 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. The p value less than <0.05 was considered as statistically significant in all the experiments; Scale bar 100 µm.

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Incubation, Staining, Concentration Assay, Fluorescence, Microscopy, Software

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced NRF2 translocation in BV2 microglial cells. The BV2 and BV2-G cells were seeded in a T25 culture flask and cultured under standard conditions. Cells were incubated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were washed with PBS lysed and cytosolic and nuclear fractions separated. Aliquots were subjected to western blot analysis (A). Decreased nuclear and increased cytosolic expression level of NRF2 was found in BV2-G cells as compared with BV2 cells. Bar graphs shows the mean densitometry analysis of bands after normalizing with β-actin as a loading control of each group (B and C). Values are given as mean ± SEM of three experiments in each group. Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. p<0.001 untreated controls vs MPP+-treated cells; p<0.05 and p<0.001 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. The p value less than <0.05 was considered as statistically significant in all the experiments.

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Translocation Assay, Cell Culture, Incubation, Western Blot, Expressing, Software

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced COX2 and NOS2 expression in BV2 microglial cells. The BV2 and BV2-G microglial cells were seeded in a T25 culture flask. The cells were incubated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were washed with PBS and cell lysates were prepared from these cells for western blot studies. Decreased expression of COX2 and NOS2 were found in BV2-G cells when compared with BV2 microglial cells (A). Bar graphs show the mean densitometry analysis of bands after normalizing with β-actin as a loading control of each group (B and C). Values are given as mean ± SEM of three experiments in each group. Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. p<0.001 and p<0.01 untreated controls vs MPP+-treated cells; p<0.01 and p<0.05 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. The p value less than <0.05 was considered as statistically significant in all the experiments.

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Expressing, Incubation, Western Blot, Software

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced HO-1 and ferritin expression in BV2 microglial cells. The BV2 and BV2-G cells were seeded in a T25 culture flask and cultured under standard conditions. The cells were incubated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were washed with PBS and prepared for western blot analysis. Decreased HO-1 and ferritin expressions were found in BV2-G cells as compared with BV2 cells (A). Bar graphs show the mean densitometry analysis of bands after normalizing with β-actin as a loading control of each group (B and C). Values are given as mean ± SEM of three experiments in each group. Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. p<0.001 untreated controls vs MPP+-treated cells; p<0.05 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. The p value less than <0.05 was considered as statistically significant in all the experiments.

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Expressing, Cell Culture, Incubation, Western Blot, Software

Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: CRISPR/Cas9 Editing of Glia Maturation Factor Regulates Mitochondrial Dynamics by Attenuation of the NRF2/HO-1 Dependent Ferritin Activation in Glial Cells

doi: 10.1007/s11481-019-09833-6

Figure Lengend Snippet: Effect of GMF on MPP+ induced HO-1 and ferritin expression in BV2 microglial cells. The BV2 and BV2-G microglial cells were seeded on poly-D-lysine coated coverslips. The cells were incubated with MPP+ (0.1 mM) for 24 h. After the incubation period, cells were washed with PBS and processed for immunocytochemistry to detect HO-(A) and ferritin (D) co-localization along with NRF2 expression. Quantitatively decreased total positive area and total average intensity of HO-1 (B and C) and ferritin (E and F) along with NRF2 translocation to nuclear site were found in BV2-G cells as compared with BV2 microglial cells. Statistical significance was assessed by independent student t-test using GraphPad InStat prism-7 software. p<0.05 untreated controls vs MPP+-treated cells; p<0.05 MPP+-treated BV2 cells vs MPP+-treated BV2-G cells. The p value less than <0.05 was considered as statistically significant in all the experiments; Scale bar 100 µm; au arbitrary units

Article Snippet: Statistical analysis All the results were expressed as mean ± SEM and statistical analysis of the data was done by using GraphPad InStat prism-7 software.

Techniques: Expressing, Incubation, Immunocytochemistry, Translocation Assay, Software