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FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to <t>DAT</t> (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.
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FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to <t>DAT</t> (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.
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FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to <t>DAT</t> (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.
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FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to <t>DAT</t> (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in <t>neurotransmitter</t> levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.
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Image Search Results


FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to DAT (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.

Journal: Journal of Biological Chemistry

Article Title: Functional Compartmentalization of the Plasma Membrane of Neurons by a Unique Acyl Chain Composition of Phospholipids

doi: 10.1074/jbc.m114.571075

Figure Lengend Snippet: FIGURE 7. Disruption of the OPPC domain affects protein segregation. PC12 cells that had been treated with NGF for 48 h were treated with 100 M RHC80267 in the bottom lane of A and F–J (blue in B–E) or with vehicle in the top lane of A and F–J (red in B–E) for 3 h. These cell preparations were analyzed as follows. A, in situ PLA1 activity assay by PED-A1 shown in green. B, remodeling assay with TMF from RHC80267-treated cells (blue) or from control cells (red). C, quantification of the remodeled products in B. The percentage of each remodeled product in TMF derived from RHC80267 treated cells (blue) or from control cells(red)isshown.D,RP-HPLCandMALDI-TOF-MSanalysisofTMFlipids(red,NGF;blue,NGFwithRHC80267).Thelipidsinfraction30oftheC8column(Fig. 2) of each sample were analyzed with MALDI-TOF-MS with 6.25 pmol of dimyristoylphosphatidylcholine as a calibration standard for quantification. E, OPPC content per protein in TMF (red, NGF; blue, NGF with RHC80267). Data are means from four independent experiments evaluated using two-tailed Student’s t test (*, p 0.02). Error bars, S.D. Immunofluorescent staining with mAb#15 (F; green), with antibody to Thy-I (G; green) and with antibody to DAT (H; red; double stainingthesamecellinG).I,dopamineuptakeassayusingafluorescentdopamineanalogshowningreen.J,immunofluorescentstainingwithantibodytoGo (green). The thick arrows and the thin arrows point to the presence and the absence of the activity, respectively, in each image. Bar, 40 m.

Article Snippet: Antibodies to Thy-1 (Millipore, MAB1406) and to DAT (Alomone Labs, AMT-003) were used in 100-fold dilution in combination with 5000-fold diluted Alexa 555-labeled antibody to rabbit IgG and Alexa 488-labeled antibody to mouse IgG.

Techniques: Disruption, In Situ, Activity Assay, Control, Derivative Assay, Two Tailed Test, Staining

(A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in neurotransmitter levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–E) PNN neuroprotective role in mitigating EBRO-induced alterations in neurotransmitter levels, including Acetylcholine (A) ; Dopamine (B) ; GABA (C) ; Glutamate (D) , and Serotonin (E) in brain homogenates. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: Additionally, neurotransmitters like dopamine [KLR0219; Krishgen Biosystem, Mumbai, India], glutamate [KLR1474; Krishgen Biosystem, Mumbai, India], serotonin [KLR0866; Krishgen Biosystem, Mumbai, India], acetylcholine [KLR0722; Krishgen Biosystem, Mumbai, India] , and GABA [KLR0102; Krishgen Biosystem, Mumbai, India] were evaluated ( ).

Techniques: Standard Deviation, Control