recombinant mouse alpha synuclein pffs  (Novus Biologicals)

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Plot of fold change (FC) of the size of α-syn-induced lamina vacuoles vs. BH-adjusted P value of two-sided nonparametric two-sample Mann–Whitney U test of genetic screen for α-syn modifiers using EGFP RNAi as control, using the Matlab function ranksum. The RNAi or overexpression (OE) with log2(FC) < −1 and P < 0.01 were labeled. Each data point was generated by the Matlab function plot(log2(FC), –log10( p ),’.’) using FC and P for each group. b , d Startle-induced climbing assay of flies incubated at 25 °C, expressing pan-neuronal α-syn either by nSyb-GAL4 > SNCA ( b ) or nSyb-QF2 > SNCA ( d ). c Western blot and quantification of relative α-syn protein levels in the head of flies incubated at 29 °C for 3 weeks, with genotypes nSyb-GAL4 > SNCA, nSyb-QF2 > SNCA , and respective controls by western blot. e , g Immunostaining of PAM cluster TH+ neurons ( e , g ) and pixel classification of TH+ pixels using Labkit ( e ) in flies nSyb-GAL4 > SNCA incubated at 25 °C for 32 days or 49 days ( e ), or in flies nSyb-QF2 > SNCA incubated at 29 °C for 21 days ( g ). Scale bar = 20 μm. f , h Quantification of PAM cluster TH+ neurons in flies with nSyb-GAL4 > SNCA ( f ) or nSyb-QF2 > SNCA ( h ). b , d Each data point indicates the mean climbing height of 1 vial containing 20–30 flies, measured repeatedly across timepoints. c Each data point and each lane indicates one pool of dissected fly brains. f , h Each data point indicates one fly. Number of vials ( n ) per group is 13, 14, 12, 14, 14, 14, 14, 14 for 8 time points in ( b ), respectively, and 12, 20, 16, 19 for 4 time points in ( d ), respectively. Number of pools of dissected fly brains ( n ) per group 3 for ( c ). Number of flies ( n ) per group is 50, 48, 57, 40, 20, 17, 50, 29 for ( e , f ); 71, 39, 35, 50, 64, 55 for ( g , h ). Data in ( c ) were analyzed using a two-sided T test. Data in ( b , d , f , h ) at each timepoint were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. The group comparisons in ( b , d ) were analyzed using two-way ANOVA followed by Tukey’s multiple comparisons test. The asterisks in ( b ) indicate the comparisons between α-syn with mino RNAi and α-syn with EGFP RNAi. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Source data are provided as a Source Data file. A single experiment with biological replicates was performed.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: MANN-WHITNEY, Control, Over Expression, Labeling, Generated, Climbing Assay, Incubation, Expressing, Western Blot, Immunostaining

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Schematic illustration of the glycolysis, TCA cycle, and de novo lipid synthesis pathways. The fold changes of metabolites are indicated in red. Green arrows indicate the de novo fatty acid synthesis and lipogenesis. Purple arrows indicate the malate-aspartate shuttle. Gray-filled boxes indicate the transporters on the mitochondria's inner membrane. b Short-chain fatty acids profiling in dissected fly brains with pan-neuronal α-syn or not, incubated at 29 °C for 3 weeks. The numbers indicate the average metabolite (pmol) per fly brain from the pooled samples of ten brains. The number of pools ( n ) per group is 1. N.D., not detected or had concentrations below the lower limit of quantification. N.A. not available. c Z-stack projections of the cortex of the optic lobe of flies incubated at 29 °C for 3 weeks, stained by BODIPY 493/503. Scale bar = 20 μm. d Quantification of mean intensity of BODIPY 493/503 for ( c ). e Z-stack projections of cortex LDs stained by BODIPY 493/503 in the optic lobe of flies incubated at 29 °C for 3 weeks. Scale bar = 20 μm. f Quantification of LDs for ( e ). Each data point indicates one fly. Number of flies ( n ) per group is 19, 16, 19, 17, 21, 19 for ( c , d ); 19, 16, 19, 17, 21, 19, 19, 21, 22, 25, 27, 24 for ( e , f ). Data in ( d ) were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. Data in ( f ) were analyzed two-sided with Brown–Forsythe and Welch’s ANOVA followed by Dunnett’s T3 multiple comparisons test. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Glu glutamate, Asp aspartate, MPC mitochondrial pyruvate carrier, CTP citrate transport protein, AGC aspartate glutamate carriers, MKA malate α-ketoglutarate antiporter, GOT1 glutamate oxaloacetate transaminase 1, GOT2 glutamate oxaloacetate transaminase 2, MDH1 malate dehydrogenase 1, MDH2 malate dehydrogenase 2. Source data are provided as a Source Data file. A single experiment with biological replicates was performed.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: Membrane, Incubation, Staining

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Predicted subcellular localization of mino , Gpat4 , CG15450 , and Gnpat using MULocDeep. b Startle-induced climbing assay of flies incubated at 25 °C, with neuronal RNAi of mino , Gpat4 , CG15450 , or Gnpat in fly brains with pan-neuronal α-syn or not. c Mean actograms of flies incubated at 25 °C in 12-h light/dark cycles for 20 consecutive days. The periods of light and dark are indicated using open or filled rectangles on the top, respectively. d Quantification of daily total locomotor activities for each fly of ( c ). f , g Immunostaining of PAM cluster TH+ neurons in flies incubated at 29 °C for 21 days. Scale bar = 20 μm. e , h Quantification of PAM cluster TH+ neurons for ( f , g ), respectively. g , h FSG67 was added to fly food at 1 mM. i Quantification of daily total locomotor activities for flies incubated at 25 °C and fed with FSG67 or not. b Each data point indicates the mean climbing height of 1 vial containing 20–30 flies, measured repeatedly across timepoints. d , i Measurements were taken from distinct flies for each timepoint, with each data point as one fly, and were measured repeatedly across timepoints. Each data point indicates one fly in ( e , h ). Number of flies ( n ) per group is 16 for ( b) ( Syb-QF2, nSyb-GAL4 ); 8 for ( b ) ( Syb-QF2, nSyb-GAL4 > gene RNAi ); 16, 8, 8 for ( b ) (day 3, 6, 12, for all the groups containing Syb-QF2 > SNCA ); 4 for ( c , d ); 46, 38, 40, 26, 39, 54, 54, 52, 49, 47 for ( e ); 65, 62, 66 for ( h ); 7 for ( i ). Data in ( b ) at each timepoint and data in ( e , h ) were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. The group comparisons in ( b , d , i ) were analyzed using two-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Source data are provided as a Source Data file. A single experiment with biological replicates was performed.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: Climbing Assay, Incubation, Immunostaining

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Western blot of α-syn, Pgi, Ldh, and pS129 in dissected fly brains of flies incubated at 29 °C for 3 weeks. b Quantification of relative protein levels in ( a ). c Western blot of α-syn higher-order oligomers formation in dissected fly brains expressing pan-neuronal α-syn, with flies incubated at 29 °C for 3 weeks. A 3D heatmap of α-syn was shown on the right. d Quantification of the ratio between α-syn oligomers and monomers, or the percentage of monomers and oligomers to total α-syn in ( c ). e α-syn flies were fed with food supplemented with 1 mM FSG67 or not, incubated at 25 °C for 3 weeks before oligomer analysis. Each data point in ( b , d , e ) and each lane in ( a , c ) indicates one pool of dissected fly brains. Number of pools of dissected fly brains ( n ) per group is 4 for α-syn and Ldh, 2 for Pgi, and 3 for pS129/α-syn in ( b ); 5 for ( d ); 4 for ( e ). Data in ( b ) were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. Data in ( d , e ) were analyzed with two-way ANOVA followed by Fisher’s LSD test. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Source data are provided as a Source Data file. A single experiment with biological replicates was performed.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: Western Blot, Incubation, Expressing

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Imaging of mitochondrial H 2 O 2 sensor in photoreceptor axons of flies incubated at 25 °C for 2 weeks, expressing H 2 O 2 sensor Rh1-GAL4 > Mito-roGFP2-Orp1 , and expressing α-syn (Rh1 > SNCA ) or not. The H 2 O 2 increased as the merged color shift from green to yellow and red. Scale bar = 5 μm. b Quantification of the ratio between mean intensity in 405 nm and 488 nm channels. Treatment with 20 mM H 2 O 2 for 5 min was used as a positive control. c , e , g Imaging of pan-neuronal MitoTimer in the cortex of optic lobe reflecting the maturation of mitochondria from green to red color with time ( c ), or at day 5 of flies incubated at 25 °C ( e ), or fed with FSG67 or not and incubated at 22 °C or 25 °C for 1 day, 1 week, and 2 weeks ( g ). Scale bar = 10 μm ( c , g ) and 5 μm ( e ). d , f , h Quantification of MitoTimer aging index using the ratio of mean intensity of red to green channels of MitoTimer in ( c , e , g ), respectively. Each data point indicates one fly. Number of flies ( n ) per group is 21, 21, 21, 27, 33, 31, 22 for ( b ); 33, 16, 20, 26, 28 for multiple time points in ( d ) ( nSyb-QF2, nSyb-GAL4 ); 27, 21, 16, 24, 29 for multiple time points in ( d ) ( nSyb-QF2, nSyb > mino RNAi ); 38, 18, 19, 29, 23 for multiple time points in ( d ) ( nSyb-QF2 > SNCA, nSyb-GAL4 ); 29, 23, 22, 33, 26 for multiple time points in ( d ) ( nSyb-QF2 > SNCA, nSyb > mino RNAi ); 32, 27, 30, 30, 29, 26, 29, 29, 57, 50, 36, 41, 48, 40 for ( f ); 31, 25, 31, 29, 33, 26; (31, 25 shared day 1 data), 31, 32, 34, 30, 31, 31, 30, 29 for ( h ). Data in ( d ) at each timepoint and data in ( b , f , h ) were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Source data are provided as a Source Data file. A single experiment with biological re p licates was performed.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: Imaging, Incubation, Expressing, Positive Control

Journal: Nature Communications

Article Title: Glycerol 3-phosphate acyltransferase exacerbates α-synuclein-induced toxicity by increasing lipid peroxidation

doi: 10.1038/s41467-026-68325-3

Figure Lengend Snippet: a Mouse primary neurons were transfected with α-syn monomers or PFF with 75 μM FSG67 for 2 weeks and stained with antibody against phosphorylated α-syn pS129 and lipid peroxidation product 4-HNE. Scale bar = 30 μm. b , c Quantifications of pS129 and 4-HNE intensity sum. For pS129 ( b ), each data point is the mean of technical replicates from one animal (a total of three independent experiments using three animals). Number of animals ( n ) per group is 3. d Mouse primary neurons were transfected with PFF with 0, 75, or 150 μM FSG67 for 2 weeks and stained with antibody against pS129 and the lipid peroxidation product MDA. Scale bar = 30 μm. e Quantifications of MDA intensity sum. For 4-HNE ( c ) and MDA ( e ), all data points are technical replicates ( > 30) from only one animal. Number of animals ( n ) per group is 1. f , g Mouse primary neurons were transfected with PFF with 0, 75, or 150 μM FSG67 for 3 weeks and live-stained with BODIPY C11 ( f ) for the lipid peroxidation analysis ( g ). Each data point is the mean of technical replicates ( > 48) from one animal (total three independent experiments using three animals). The number of animals ( n ) per group is 3. Scale bar = 40 μm. Data were analyzed with one-way ANOVA followed by Tukey’s and Dunnett’s multiple comparisons test. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Source data are provided as a Source Data file.

Article Snippet: At day 7, mice primary cortical neurons were treated with 1 μg/ml α-syn monomers (SPR-323, StressMarq) or 1 μg/ml synuclein PFF (SPR-322, StressMarq) with or without 75 μM or 150 μM FSG (HY−112489, MedChemExpress LLC) for 2 weeks.

Techniques: Transfection, Staining