Journal: Molecular Neurodegeneration

Article Title: VCP regulates early tau seed amplification via specific cofactors

doi: 10.1186/s13024-024-00783-z

Figure Lengend Snippet: List of VCP cofactors and their proposed functions

Article Snippet: Nsfl1c Polyclonal Antibody , Fisher Scientific , PA5-21,633.

Techniques: Membrane, Ubiquitin Proteomics

Journal: Molecular Neurodegeneration

Article Title: VCP regulates early tau seed amplification via specific cofactors

doi: 10.1186/s13024-024-00783-z

Figure Lengend Snippet: VCP cofactors differentially regulate tau seeding. VCP cofactors were either knocked out via CRISPR/Cas9 ( A-D ) or knocked down via siRNA ( E–G ) in v2L biosensors prior to exposure to increasing amounts of tau fibrils. A Knockout of FAF2 increased tau seeding whereas knockout of B ATXN3, C NSFL1C, and D UBE4B reduced tau seeding. P values: FAF2 (*** 0.0001, **** < 0.0001); ATXN3 (**** < 0.0001); NSFL1C (*** 0.0002); UBE4B (**** < 0.0001). E Knockdown of NGLY1, F NPLOC4, and G OTUB1, decreased tau seeding. P values: NGLY1(**** < 0.0001); NPLOC4 (**** < 0.0001); OTUB1 (*** 0.0004, **** < 0.0001, *** 0.0001). Graphs are representative of n = 3 independent experiments, with each data point derived from technical triplicate. Error bars represent S.D. Some error bars are too small to be visible. H Cofactor KO did not affect tau uptake. P values: ns = 0.9819, 0.9988, 0.9956, 0.9928, in order of bars on the graph. I Cofactor KD did not affect tau uptake. P values: ns = 0.9795, 0.1856, 0.3928, in order of bars on the graph. Error bars represent S.E.M ( n = 3). One-Way ANOVA with a 95% confidence interval

Article Snippet: Nsfl1c Polyclonal Antibody , Fisher Scientific , PA5-21,633.

Techniques: CRISPR, Knock-Out, Knockdown, Derivative Assay

Journal: Molecular Neurodegeneration

Article Title: VCP regulates early tau seed amplification via specific cofactors

doi: 10.1186/s13024-024-00783-z

Figure Lengend Snippet: List of Reagents

Article Snippet: Nsfl1c Polyclonal Antibody , Fisher Scientific , PA5-21,633.

Techniques: Protease Inhibitor, Cell Culture, Transfection, Magnetic Beads, Sequencing, Modification

Journal: Frontiers in Neuroscience

Article Title: Behavioral benefits of GSK-3β inhibition and state-dependent microtubule signatures in the Fmr1 -KO mouse

doi: 10.3389/fnins.2025.1643439

Figure Lengend Snippet: Experimental design (see also main text). Three experimental cohorts were generated for this study. (A) Behavioral and α -tubulin PTMs phenotyping - A cohort of 3-month-old male mice, WT and Fmr1 -KO, was used to assess baseline behavioral and α -tubulin PTMs phenotypes. Behavioral assays included the open field, elevated plus maze, novel object recognition, marble burying, and 3-chamber sociability test. Mice were sacrificed 24 h after the last behavioral test for molecular analysis of hippocampal tissue. (B) Molecular phenotyping - A separate mixed-sex cohort was used to investigate sex-dependent molecular changes in hippocampal lysates. The cohort included five genotypes: WT males, Fmr1 -KO males, WT females, Fmr1 -heterozygous females, and Fmr1 -KO females. Molecular analysis included: α -tubulin PTMs, FMRP and synaptic markers. (C) Effects of GSK-3β inhibitors – A third of 3-month-old male mice, WT and Fmr1 -KO, was used to evaluate the effects of GSK-3β inhibition on behavioral and molecular endpoints. WT and Fmr1 -KO mice received daily intraperitoneal (i.p.) injections of either SB216763 (3 mg/kg) or AF3581 (10 mg/kg) for 10 consecutive days. On Day 9, mice were tested in the marble burying task, and on Day 10 in the 3-chamber sociability test, performed 30 min post-treatment. Mice were sacrificed 24 h after the final injection for molecular analysis of hippocampal tissue.

Article Snippet: Glu-Tub was detected using anti α -tubulin rabbit monoclonal antibody [clone; RM444], detyrosinated (Fisher Scientific; 17,846,771) at 1:500 overnight at 4°C.

Techniques: Generated, Inhibition, Injection

Journal: Frontiers in Neuroscience

Article Title: Behavioral benefits of GSK-3β inhibition and state-dependent microtubule signatures in the Fmr1 -KO mouse

doi: 10.3389/fnins.2025.1643439

Figure Lengend Snippet: Fmr1 -KO mice show significantly altered microtubule dynamics in the hippocampus. (A) male Fmr1-KO mice ( n = 10) had significantly higher Acet/Tot-Tub α -tubulin ratios compared to WT ( n = 10), and (B) significantly lower Tyr/Glu-Tub α -tubulin ratios. There were also sex-dependent effects for both Acet/Tot-Tub and Tyr/Glu-Tub α -tubulin. (C) Female WT ( n = 9) and Fmr1 -KO ( n = 10) mice had significantly lower Acet/Tot-Tub compared to their respective genotype-matched males [WT ( n = 11) and Fmr1 -KO ( n = 13) mice] while heterozygous females ( n = 7) had similar Acet/Tot-Tub α -tubulin ratios to WT males. (D) Tyr/Glu-Tub levels are significantly higher in female WT ( n = 8) and Fmr1 -KO ( n = 11) mice compared to their respective genotype-matched males [WT ( n = 11) and Fmr1 -KO ( n = 13) mice], but Fmr1 -KO mice of both sexes show significant reductions in Tyr/Glu ratios compared to WTs. Heterozygous females ( n = 7) were not significantly different to WT males. (E) Complete KO of the Fmr1 gene prevents FMRP expression as expected but heterozygous mice produce approximately 73% of WT FMRP levels [male: WT ( n = 11) and Fmr1 -KO ( n = 13)] mice; [female: WT ( n = 9), Fmr1 -KO ( n = 10) and Fmr1 -heterozygous ( n = 7)]. (F–H) Expression of synaptic markers [male: WT ( n = 11) and Fmr1 -KO ( n = 13)] mice; [female: WT ( n = 9), Fmr1 -KO ( n = 10) and Fmr1 -heterozygous ( n = 7)]. (F) The postsynaptic protein spinophilin is significantly increased in Fmr1 -KO mice but produced at levels similar to WT by Fmr1 -heterozygous females. There are no significant differences in (G) the presynaptic protein synaptophysin or (H) the postsynaptic protein PSD-95. Data are expressed as MEAN ± SEM. (A,B) Unpaired T -test, ** p < 0.01; *** p < 0.001 versus WT controls. (C–H) Two-way ANOVA with sex (male, female) and genotype (WT, heterozygous, KO) as factors, * p < 0.05; ** p < 0.01; **** p < 0.0001 versus male WT controls. + p < 0.05; ++++ p < 0.0001 versus WT females. # p < 0.05; #### p < 0.0001 versus Fmr1 -KO females.

Article Snippet: Glu-Tub was detected using anti α -tubulin rabbit monoclonal antibody [clone; RM444], detyrosinated (Fisher Scientific; 17,846,771) at 1:500 overnight at 4°C.

Techniques: Expressing, Produced

Journal: Frontiers in Neuroscience

Article Title: Behavioral benefits of GSK-3β inhibition and state-dependent microtubule signatures in the Fmr1 -KO mouse

doi: 10.3389/fnins.2025.1643439

Figure Lengend Snippet: Effects of GSK-3β inhibition on behavioral and molecular endpoints in Fmr1 -KO mice. (A–D) Behavioral readouts [WT: Vehicle ( n = 10), SB216763( n = 10) and AF3581 ( n = 10); Fmr1 -KO: Vehicle ( n = 10), SB216763 ( n = 10) and AF3581 ( n = 10)]. (A) Perseverative behavior was rescued by GSK-3β inhibition as marble burying was significantly reduced in Fmr1 -KO mice following treatment with SB216763 and AF3581. (B) Vehicle-treated Fmr1 -KO mice were significantly impaired in the social discrimination trial of the 3-chamber sociability test compared to WT, this was significantly rescued by SB216763 and partially by AF3581. (C) Social recognition was not different in Fmr1 -KO mice nor did GSK-3β inhibition have any effect on this behavior. (D) Overall, Fmr1 -KO mice showed significantly higher levels of locomotor activity compared to WT mice. Only vehicle-treated Fmr1 -KO mice showed a specific hyperlocomotion, Fmr1 -KO mice treated with either SB216763 or AF3581 were not significantly different to WT mice at any time point. (E) All three Fmr1 -KO treatment groups had higher Acet/Tot-Tub compared to vehicle-treated WT mice, with both SB216763- and AF3581-treated Fmr1 -KO mice showing significantly higher expression. (F) There were no significant differences between Tyr/Glu α -tubulin ratios in any of the treatment groups, though SB216763-treated Fmr1 -KO mice were trending toward an increase compared to WT and Fmr1 -KO vehicle controls. Data are expressed as MEAN ± SEM. (A-C) Two-way ANOVA with genotype (WT, Fmr1 -KO) and treatment (vehicle, SB216763, AF3581) as factors, * p < 0.05; ** p < 0.01 versus vehicle-treated Fmr1 -KO mice. (D) Three-way repeated measures ANOVA with genotype (WT, Fmr1 -KO) and treatment (vehicle, SB216763, AF3581) as between-subjects factors, and trial (habituation, Trial 1 and Trial 2) as a within-subjects factor, * p < 0.05; ** p < 0.01 versus vehicle-treated Fmr1 -KO mice. (E,F) One-way ANOVA with treatment groups (vehicle, SB216763, AF3581) as the between-subjects factor, ** p < 0.01 versus vehicle-treated WT mice.

Article Snippet: Glu-Tub was detected using anti α -tubulin rabbit monoclonal antibody [clone; RM444], detyrosinated (Fisher Scientific; 17,846,771) at 1:500 overnight at 4°C.

Techniques: Inhibition, Activity Assay, Expressing

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-CD44 (rabbit monoclonal) , Fisher Scientific , Cat. #:15266957, RRID:N/A , Blocking experiment (10 μg/mL).

Techniques: Blocking Assay, Recombinant

Journal: bioRxiv

Article Title: Translational toolkit for reproducible, cross-study profiling of human ageing hallmarks in human blood and tissue

doi: 10.64898/2026.04.20.719545

Figure Lengend Snippet: A. Experimental design for assessing nutrient sensing (mTOR activation) and autophagic flux in cryopreserved PBMCs using flow cytometry. B. Representative histograms showing phospho-S6 expression in PBMCs at baseline (untreated) and following 30-minute stimulation with PMA (phorbol-12-myristate-13-acetate) and ionomycin. C. Representative histogram showing LC3-II (microtubule-associated protein 1A/1B-light chain 3) accumulation in autophagosomes of T cells at baseline and after 2-hour treatment with 100 µM chloroquine (Cq). Radar plots depicting phospho-S6 expression assessed as median fluorescence intensity (MFI) in (D) CD4+ and (E) CD8+ T cells at baseline and after 30-minute stimulation with PMA and ionomycin in young and aged participants. F. Radar plots showing phospho-S6 fold-change relative to young participants in CD4+ and CD8+ T cells following stimulation. G. Baseline autophagosomal LC3-II expression in T cells from young (blue, n=6) and aged (red, n=12) participants. H. T-cell autophagic flux following 2-hour treatment with 100 µM chloroquine. Data are mean ± S.E.M. Statistical significance was assessed (D-H) unpaired t-tests.

Article Snippet: Cells were then washed with 100 μl of 1X permeabilization buffer (5 minutes, 250 x g, at 4 °C) and resuspended in 50 μl of permeabilization buffer with 1 μl of PE Phospho-S6 antibody (Fisher Scientific, cat. no.: 12-9007-42, clone: cupk43k) for 20 min on ice in the dark.

Techniques: Activation Assay, Flow Cytometry, Expressing, Fluorescence