Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: LRP1 expression is increased in the nigrostriatal system of a monkey model of PD induced by α-syn PFFs injection. (A) Clinical rating score of monkeys after stereotactic injection of 600 μg of α-syn PFFs or an equal volume of normal saline for 4 months. (B) Immunohistochemistry detection of TH in the STR (left) and SN (right). The TH signal intensity in the STR and the number of TH-positive neurons in the SN were lower in the PFF group. The black arrow heads indicate typical TH-positive neurons. Scale bars: 100 μm. (C) Quantitative immunohistochemistry density analysis of TH in the STR. (D) Quantitation of the ratio of TH-positive neurons in the SN in the PFF group compared with the Sham group. (E) Western blot analysis of TH, LRP1, and α-syn expression levels in the monkey STR. (F–H) Densitometric analysis of TH (F), LRP1 (G), and α-syn (H) expression levels in the STR. (I) Western blot analysis of TH, LRP1, and α-syn expression levels in the monkey SN. (J–L) Densitometric analysis of TH (J), LRP1 (K), and α-syn (L) expression levels in the SN. Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. Sham group (unpaired t -test). LRP1: Low-density lipoprotein receptor-related protein 1; PD: Parkinson’s disease; PFF: pre-formed fibril; SN: substantia nigra; STR: striatum; TH: tyrosine hydroxylase; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Expressing, Injection, Saline, Immunohistochemistry, Quantitation Assay, Western Blot

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: LRP1 expression is increased in the nigrostriatal system of a mouse model of PD induced by injection with α-Syn PFFs. (A) Visual gait analysis of walking, gait, and footprint pressure in mice after stereotactic injection with 5 μg of α-Syn PFFs or an equal volume of PBS for 4 weeks ( n = 6). (B) Normal step sequence in mice ( n = 6). (C) Average speed in mice ( n = 6). (D) Tripod support time in mice ( n = 6). (E) Immunohistochemistry staining for TH in the STR. The images in the second row are enlarged images of the areas indicated in black boxes in the upper row. The TH signal intensity in the STR was lower in the PFF group. Scale bars: 200 μm, 40 μm (enlarged). (F) Immunohistochemistry staining for TH in the SN. The images in the second row are enlarged images of the areas indicated in black boxes in the upper row, and the white arrowheads indicate typical TH-positive neurons. There were fewer TH-positive neurons in the SN in the PFF group. Scale bars: 100 μm, 20 μm (enlarged). (G) Quantitative immunohistochemistry density analysis of TH in the STR, compared with the Sham group ( n = 3). (H) Quantitation of the ratio of TH-positive neurons in the SN in the PFF group compared with the Sham group ( n = 3). (I) Western blot analysis of TH, LRP1, and α-Syn expression levels in the mouse STR after injection with 5 μg of α-Syn PFFs or an equal volume of PBS ( n = 3). (J–L) Densitometric analysis of TH (J), LRP1 (K), and α-Syn (L) expression levels in the STR. (M) Western blot analysis of TH, LRP1, and α-Syn expression levels in the mouse SN ( n = 3). (N–P) Densitometric analysis of TH (N), LRP1 (O), and α-Syn (P) expression levels in the SN ( n = 3). Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . Sham group (unpaired t -test). LRP1: Low-density lipoprotein receptor-related protein 1; PBS: phosphate buffered solution; PD: Parkinson’s disease; PFF: pre-formed fibril; SN: substantia nigra; STR: striatum; TH: tyrosine hydroxylase; α-Syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Expressing, Injection, Sequencing, Immunohistochemistry, Staining, Quantitation Assay, Western Blot

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: Exogenous α-syn PFFs upregulate LRP1 expression in PC12 cells. (A) Viability of PC12 cells incubated with different doses (0, 5, 10, 20, or 50 µg/mL) of α-Syn PFFs for 24 hours. ** P < 0.01, *** P < 0.001, vs . 0 µg/mL group. (B) Western blot analysis of LRP1 and α-syn expression levels in PC12 cells. (C, D) Densitometric analysis of LRP1 (C) and α-syn (D) expression levels. (E) Immunofluorescence analysis of LRP1 (green, Alexa Fluor 488), α-syn (red, Alexa Fluor 594), and DAPI (blue) expression. The LRP1 and α-syn signal intensities were stronger in PFF group than in Con group. Scale bars: 40 μm. (F, G) Quantitative immunofluorescence intensity analysis of LRP1 (F) and α-syn (G). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . Con group; # P < 0.05, ## P < 0.01, vs. monomer group. Data are expressed as mean ± SD ( n = 3). Con: Control; DAPI: 4′,6-diamidino-2-phenylindole; PFF: pre-formed fibril; LRP1: low-density lipoprotein receptor-related protein 1; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Expressing, Incubation, Western Blot, Immunofluorescence, Control

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: LRP1 knockdown rescues the dopaminergic damage induced by exogenous α-syn PFFs. (A, B) Western blot analysis (A) and densitometric analysis (B) of TH expression levels in the STR of mice treated with PFFs. (C, D) Western blot analysis (C) and densitometric analysis (D) of TH expression levels in the SN of mice treated with PFFs. (E) Immunohistochemistry staining for TH in the STR of mice treated with PFFs and LRP1 siRNA. The decrease in relative TH intensity in STR observed in the PFF group was rescued by LRP1 siRNA treatment. The images in the second row are enlarged images of the areas indicated by black boxes in the upper row. Scale bars: 200 μm, 40 μm (enlarged). (F) Quantitative immunohistochemistry density analysis of TH in the STR. (G) Immunohistochemistry analysis of TH in the SN of mice treated with PFFs. The decreased ratio of TH-positive neurons in the SN of the Scramble + PFF group (PFF group) was rescued by LRP1 siRNA treatment. The images in the second row are enlarged images of the areas indicated by black boxes in the upper row, and the white arrowheads indicate typical TH-positive neurons. Scale bars: 100 μm, 20 μm (enlarged). (H) Quantitation of the ratio of TH-positive neurons in the SN compared with the PBS group. Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, vs. Scramble + PBS group (PBS group); # P < 0.05, ## P < 0.01, vs. scramble + PFF group (PFF group). LRP1: Low-density lipoprotein receptor-related protein 1; PBS: phosphate buffered solution; PFF: pre-formed fibril; siRNA: small interfering RNA; SN: substantia nigra; STR: striatum; TH: tyrosine hydroxylase; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Knockdown, Western Blot, Expressing, Immunohistochemistry, Staining, Quantitation Assay, Small Interfering RNA

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: LRP1 suppression reduces the transmission of exogenous α-syn in vivo . (A) Immunofluorescence staining for LRP1 (green, Alexa Fluor 488), α-syn (red, Alexa Fluor 594), and DAPI (blue) in the mouse STR after stereotactic injection with α-syn PFFs for 6 weeks with or without LRP1 knockdown. The increase in the relative fluorescence intensity of LRP1 and α-syn in the STR of PFFs-treated mice was rescued by LRP1 siRNA treatment. Scale bars: 100 μm. (B, C) Quantitative immunofluorescence intensity of LRP1 (B) and α-syn (C) in mouse STR. (D) Immunofluorescence staining for LRP1 (green, Alexa Fluor 488), α-syn (red, Alexa Fluor 594), and DAPI (blue) in the SN of mice after stereotactic injection with α-syn PFFs for 6 weeks with or without LRP1 knockdown. The increase in the relative fluorescence intensity of LRP1 and α-syn in the SN of mice treated with PFFs was rescued by LRP1 siRNA treatment. The white arrowheads indicate typical cells exhibiting α-syn and LRP1 expression. Scale bars: 100 μm. (E, F) Quantitative immunofluorescence intensity of LRP1 (E) and α-syn (F) in the mouse SN. (G) Western blot analysis of LRP1 and α-syn expression levels in the mouse STR. (H, I) Densitometric analysis of LRP1 (H) and α-syn (I) expression levels in the mouse STR. (J) Western blot analysis of LRP1 and α-syn expression levels in the mouse SN. (K, L) Densitometric analysis of LRP1 (K) and α-syn (L) expression levels in the mouse SN. Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, vs. Scramble + PBS group (PBS group); # P < 0.05, ## P < 0.01, vs . Scramble + PFF group (PFF group). DAPI: 4′,6-Diamidino-2-phenylindole; LRP1: low-density lipoprotein receptor-related protein 1; PBS: phosphate buffered solution; PFF: pre-formed fibril; siRNA: small interfering RNA; SN: substantia nigra; STR: striatum; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Transmission Assay, In Vivo, Immunofluorescence, Staining, Injection, Knockdown, Fluorescence, Expressing, Western Blot, Small Interfering RNA

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: LRP1 mediates the uptake of α-syn PFFs by PC12 cells. (A) Western blot analysis of LRP1 and α-syn expression levels in PC12 cells with or without LRP1 knockdown (10 nM siRNA) after incubation with 10 µg/mL α-syn PFFs for 24 hours. (B, C) Densitometric analysis of LRP1 (B) and α-syn (C) expression. (D) Immunofluorescence staining for LRP1 (green, Alexa Fluor 488), α-syn (red, Alexa Fluor 594), and DAPI (blue). The increase in the relative fluorescence intensity levels of LRP1 and α-syn in the PFF group was rescued by LRP1 siRNA treatment. Scale bars: 40 μm. (E, F) Quantitative immunofluorescence intensity analysis of LRP1 (E) and α-syn (F). Data are expressed as mean ± SD ( n = 3). ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . Con group; # P < 0.05, ## P < 0.01, #### P < 0.0001, vs . PFF/– group. Con: Control; DAPI: 4′, 6-diamidino-2-phenylindole; PFF: pre-formed fibril; LRP1: low-density lipoprotein receptor-related protein 1; siRNA: small interfering RNA; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Western Blot, Expressing, Knockdown, Incubation, Immunofluorescence, Staining, Fluorescence, Control, Small Interfering RNA

Journal: Neural Regeneration Research

Article Title: Low-density lipoprotein receptor–related protein 1 mediates α-synuclein transmission from the striatum to the substantia nigra in animal models of Parkinson’s disease

doi: 10.4103/NRR.NRR-D-23-01965

Figure Lengend Snippet: Lysine residues in the α-syn N-terminus are vital for LRP1-mediated α-Syn internalization. (A) Heatmap of amino acids in different α-syn domains. (B) Western blot analysis of LRP1 and α-Syn levels in PC12 cells after addition of α-syn PFFs (10 µg/mL) with or without capping of lysine residues for 24 hours. (C, D) Densitometric analysis of LRP1 (C) and α-syn (D) expression levels. (E) Immunofluorescence staining for LRP1 (green, Alexa Fluor 488), α-Syn (red, Alexa Fluor 594), and DAPI (blue). The increase in the relative fluorescence intensity of LRP1 and α-syn in the PFF group was rescued by lysine capping of α-syn. Scale bars: 40 μm. (F, G) Quantitative immunofluorescence intensity analysis of LRP1 (F) and α-syn (G). Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, **** P < 0.0001, vs. Con group; # P < 0.05, ## P < 0.01, vs . PFF group. Con: Control; DAPI: 4′,6-diamidino-2-phenylindole; LRP1: low-density lipoprotein receptor-related protein 1; NHS: sulfo-NHS acetate; PFF: pre-formed fibril; α-syn: α-synuclein.

Article Snippet: α-Syn PFFs (SPR-322, StressMarq, Victoria, Canada) were sonicated with a handheld probe (VCX130, SONICS & MATERIALS, Newtown, CT, USA) as follows: amplitude 30% and time 15 seconds (pulse on 3 seconds, pulse off 6 seconds) (Volpicelli-Daley et al., 2014; Sun et al., 2022b).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Fluorescence, 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 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