Journal: PLOS Neglected Tropical Diseases

Article Title: Plasmodium knowlesi tryptophan-rich antigen 40.1 binds human erythrocyte spectrin alpha

doi: 10.1371/journal.pntd.0014700

Figure Lengend Snippet: ( A) Protein solubility analysis of PkTRAg40.1 in P. knowlesi A1-H.1-infected erythrocytes. After hypotonic release of soluble proteins (Hypo), the hypotonic insoluble fractions were sequentially treated with sodium carbonate (Na 2 CO 3 ), 5M urea, and 1% Triton X-100. Supernatant (S) and pellet (P) fractions from each extraction step were collected. (B) Protease protection assay of PkTRAg40.1. Infected erythrocytes were treated with PBS, streptolysin O (SLO), or saponin in the presence (+) or absence (−) of proteinase K (pK). Insoluble fractions were subsequently prepared. PBS preserves both the erythrocyte and parasitophorous vacuole membranes (PVM), SLO selectively permeabilizes the erythrocyte membrane, and saponin disrupts the PVM while largely preserving the parasite plasma membrane. For both assays, samples were analyzed by immunoblotting using anti-PkSBP1 rabbit sera, anti-PkTRAg40.1 mouse sera, anti-spectrin alpha polyclonal antibody, and anti-aldolase polyclonal antibody. Red arrowheads indicate the expected ~ 40 kDa PkTRAg40.1 band, whereas green arrowheads indicate additional higher-molecular-weight immunoreactive bands consistently detected across the assays. Uncropped western blots are shown in and .

Article Snippet: Slides were incubated with mouse anti-PkTRAg40.1 (1:100), rabbit anti-PkSBP1 (1:100), or rabbit anti-spectrin alpha (1:200, Abclonal) antibodies at 37°C for 1 h. Secondary antibodies used were Alexa Fluor 488-conjugated goat anti-mouse IgG and Alexa Fluor 594-conjugated goat anti-rabbit IgG (1:500, Invitrogen).

Techniques: Solubility, Infection, Extraction, Membrane, Preserving, Clinical Proteomics, Western Blot, Molecular Weight

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, Schematic of βII and βIII spectrins expression in the somatodendritic compartment, axon initial segment (AIS) and distal unmyelinated axon. b, Comparison of βII and βIII spectrins sequences and binding domains. c, d, e, Representative confocal images for βII-spectrin (orange), βIII-spectrin (magenta), CAAX-GFP (grey) and Homer1C (cyan) in a filopodium (a), a mature spine (b) or a dendrite (c) at 21 DIV. Scale bars, 1 µm for filopodium and spine, 2 µm for dendrite. Note that the display contrast was purposely kept constant. f, g, Fluorescence intensity of βII-spectrin and βIII-spectrin in spine necks and dendrites at 10 DIV and 21 DIV normalized to the average intensity of filopodia at the same time-point. Each box shows the median ± percentile. n between 98 and 160 spines or 84 and 97 dendrites from 20 to 30 neurons examined over 3 independent experiments for each time point. Mann-Whitney test, *: p < 0.05, ***: p < 0.001, ****: p < 0.0001. h, i, Correlation scatter plots between βII-spectrin and βIII-spectrin normalized fluorescence intensities in spine neck or dendrite at 10 DIV and 21 DIV. n between 98 and 160 spines or 84 and 97 dendrites from 20 to 30 neurons examined over 3 independent experiments for each time point. r , Pearson’s r coefficient; P, P value.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Expressing, Comparison, Binding Assay, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, b, c, Representative d STORM images of βII-spectrin and its 1D autocorrelation amplitude in axons (a), dendrites (b), and spine necks with PSD95 labelling observed in epifluorescence (c). The non-periodic control was obtained by measuring the 1D autocorrelation amplitude of a cytosolic mCherry labelling in axons, dendrites or spine necks. n between 45 and 80 regions of interest examined over at least 3 independent experiments. d, Spacing between βII-spectrin epitopes. e, f, Representative d STORM images of βIII-spectrin and its 1D autocorrelation amplitude in dendrites (e) and spine necks (f). n between 45 and 80 regions of interest examined over at least 3 independent experiments. g, Spacing between βIII-spectrin epitopes. h, i, Representative two-color STORM images of βII-spectrin and βIII-spectrin and autocorrelation (AC) or cross-correlation amplitudes in dendrites (h) and spine necks with Homer1C labelling (i). n = 76 regions of interest examined over 3 independent experiments. j, Spacing between β-spectrin epitopes after merging the two channels. Each box shows the median ± percentile. Scale bars, 1 µm. Mann-Whitney test, ns: non-significant, ****: p < 0.0001.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Control, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, Schematic of βII-, βIII- and ɑII-spectrin in the dendrite. Arrowheads correspond to the same area. b , Quantification of the distances between clusters of localizations on XZ projections. c-f , Two-color 3D MINFLUX of βII- and βIII-spectrins in dendrite (c and d) and dendritic spine neck (e and f). Scale bars, 500 nm in c and e, 100 nm in d and f. Arrowheads highlight clusters of localizations that appear in pairs, independently of the β-spectrin isoform, mostly visible in dendrites. (d) and (f) XZ projections of regions in (c) and (e) respectively, show discrete clusters regularly spaced independently of the β-spectrin paralogue, suggesting that βII- and βIII-spectrins are radially periodic.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques:

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, b, c, Representative d STORM images of βII-spectrin in a βIII-spectrin KO axon (a), dendrite (b) and spine (c), followed by βII-spectrin localization intensities, 1D autocorrelation amplitudes and spacing measured in the associated neurites. The non-periodic controls are the same as in . PSD95 was imaged using epifluorescence microscopy. n between 30 and 88 regions of interest per neurite examined over 5 independent experiments. d, e, Representative d STORM images of βIII-spectrin in a βII-spectrin KO dendrite (d) and spine (e), followed by βIII-spectrin localizations intensities, 1D autocorrelation amplitudes and spacing measured in the associated neurites. n between 30 and 88 regions of interest per neurite examined over at least 3 independent experiments. Each box represents the median with percentile. Scale bars, 1 µm. Mann-Whitney tests, ns: non-significant, *: p < 0.05, ****: p < 0.0001.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Epifluorescence Microscopy, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, b, c, Representative d STORM images of ɑII-spectrin in control or KOs axons (a), dendrites (b) and spines (c), followed by ɑII-spectrin mean intensities, 1D autocorrelation amplitudes and spacing measured in the corresponding compartments. Only conditions where ɑII-spectrin mean intensity median value was above one-quarter of the controls ɑ2-spectrin values were analysed for autocorrelation and spacing. The non-periodic controls are the same as . PSD95 was imaged using epifluorescence microscopy. n between 14 and 53 regions of interest per neurite examined over 3 independent experiments. Each box represents the median with percentile. Scale bars, 1 µm. Mann-Whitney tests, ns: non-significant, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Control, Epifluorescence Microscopy, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, Scheme of βII- and βIII-spectrins highlighting the mutations disrupting interaction with phosphoinositides, ankyrins or actin. b, c, Representative spinning disk images of neurons re-expressing βII- (a) or βIII-spectrins (b) WT and mutant all tagged with eGFP, and expressing an intrabody labelling PSD95 . Scale bars in top images, 20 µm. Scale bars in bottom images, 1 µm. d, e, Ratios of the mean fluorescence intensity measured in axons, dendrites and spines in endogenous and re-expression conditions. n between 9 and 22 neurons obtained from 3 to 5 independent experiments for βII-spectrin, n between 7 and 16 neurons obtained from 3 to 4 independent experiments for βIII-spectrin. f, g, Mobile fractions of βII/III-spectrins measured 180 seconds after photobleaching. Each box represents the median with percentile. n between 12 and 24 regions of interest per neurite obtained from 3 to 5 independent experiments for βII-spectrin, n between 13 and 18 regions of interest per neurite obtained from 3 to 4 independent experiments for βIII-spectrin. Each box represents the median with percentile. Mann-Whitney tests, ns: non-significant, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Expressing, Mutagenesis, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, b Representative d STORM images of wild-type and mutant βII-spectrins (a) and wild-type and mutant βIII-spectrins (b) imaged in re-expression condition in the indicated neuronal compartments. The non-periodic controls are the same as in . PSD95 was imaged using epifluorescence microscopy. Scale bars, 1 µm. c, d βII-spectrin (a) and βIII-spectrins (b) 1D autocorrelation amplitudes measured in the indicated compartments. n between 31 and 162 regions of interest per compartment obtained from 3 to 5 independent experiments for βII-spectrin. n between 73 and 213 regions of interest per compartment obtained from 3 to 4 independent experiments βIII-spectrins. Each box represents the median with percentile. Mann-Whitney tests, ns: non-significant, *: p < 0.05, **: p < 0.01, ****: p < 0.0001. e, f, Spacing between βII-spectrin (e) or βIII-spectrin (f) epitopes obtained from the same datasets as in (c) and (d).

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Mutagenesis, Expressing, Epifluorescence Microscopy, MANN-WHITNEY

Journal: bioRxiv

Article Title: βII and βIII spectrin paralogues define robustness and specialization of the neuronal membrane periodic skeleton

doi: 10.64898/2026.03.23.713115

Figure Lengend Snippet: a, Schematic of the periodic actin-spectrin cytoskeleton in axons (βII-spectrin mostly), dendrites and dendritic spines (random βII and βIII spectrins distribution with the presence of sparse βII/βIII heterotypic tetramers). Spectrin signals are excluded from post-synaptic densities. b, Schematic of βII- and βIII-spectrins sensitivity to their binding partners.

Article Snippet: The following primary antibodies were used: βII-spectrin (BD Biosciences, 612 563, mouse IgG1 monoclonal, 1/500), βIII-spectrin (Novus Biotechnologies, NB110 58346, rabbit polyclonal, 1/500 or Santa Cruz, sc515737, monoclonal, 1/100), αII-spectrin (Biolegend, 803206, mouse IgG2b monoclonal, 1/1000), α-Adducin (Abcam, 51130, rabbit IgG polyclonal, 1/200), GFP (Avès, GFP-1020, chicken monoclonal, 1/2000), DsRed (Takara, 632496, rabbit polyclonal, 1/2000), PSD-95 (ThermoFisher, MA1-046, mouse IgG1 monoclonal, 1/500), Homer 1C (Synaptic Systems, 160004, guinea pig polyclonal, 1/500).

Techniques: Binding Assay

Journal: Journal of Functional Biomaterials

Article Title: A Borophosphate Glass Doped with Cobalt Oxide Improves Skeletal Muscle Structure and Function in Myopathic Mice

doi: 10.3390/jfb17030155

Figure Lengend Snippet: CoO-TRIM increased myofibers with centrally nucleated myofibers (CLN) and modulated proteolytic activity. ( A ) Representative images of TA muscle cross-sections at 14 dpt. Laminin (white): basal laminae; DAPI (blue): nuclei. Yellow arrows depicting position of central nuclei. ( B ) Summary values ( n = 4–5/group) for total myofibers with CLN normalized to TA muscle cross-section area (mm 2 ) at 14 dpt. ( C ) Summary values ( n = 4–5/group) for total myofibers (CLN + + CLN − ) normalized to TA muscle cross-section (mm 2 ) at 14 dpt. ( D ) Representative immunoblot for αII-Spectrin. The 145 kDa cleavage byproduct was normalized to total protein per lane, represented by the 40 kDa band from the total protein stain, and analyzed as a ratio of the 250 kDa band of αII-Spectrin. Mean densitometric data revealed a significant reduction in cleaved αII-Spectrin abundance in TA muscles of D2. mdx TRIM mice at 14 dpt. ( E ) Representative immunoblot of LC3B II and mean densitometric data at 14 dpt revealed reduced autophagosome number following CoO-TRIM administration in D2. mdx mice. ( n = 5/group); Summary values are means ± SEM. Comparisons made vs. WT and vehicle controls by 2-Way ANOVA, p < 0.05 = significant. Scale bars = 100 µm.

Article Snippet: Primary antibodies of interest were; Alpha II spectrin (RRID: AB_2194351, Cat.# sc-48382; Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:250 incubated overnight at 4 °C then 3 h at room temperature secondary concentration 1:5000) and LC3B (RRID: AB_915950, Cat.# 2775; Cell Signaling Technology, Danvers, MA, USA, secondary concentration 1:5000).

Techniques: Activity Assay, Western Blot, Staining, Muscles

Journal: Journal of Functional Biomaterials

Article Title: A Borophosphate Glass Doped with Cobalt Oxide Improves Skeletal Muscle Structure and Function in Myopathic Mice

doi: 10.3390/jfb17030155

Figure Lengend Snippet: Comparison of CLN in myofibers of WT and D2. mdx treated with Saline and TRIM mice. ( A , E ) Representative images of TA muscle cross-sections at 70 dpt and 140 dpt. Laminin (white): basal laminae; DAPI (blue): nuclei. Yellow arrows identify position of central nuclei. ( B , F ) Summary values for total myofibers with CLN (Top) and total myofibers (CLN + + CLN − ) (Bottom) normalized to TA muscle cross-section area (mm 2 ). Summary values presented for ( B ) 70 dpt and ( F ) 140 dpt. ( n = 7–8/group); scale bars = 100 µm. Proteolytic activity is altered following CoO-TRIM treatment. ( C , G ) Representative immunoblots for αII-Spectrin. The 145 kDa cleavage byproduct was normalized to total protein per lane, represented by the 40 kDa band from the total protein stain, and analyzed as a ratio of the 250 kDa band of αII-Spectrin. Mean densitometric data revealed a significant reduction in cleaved αII-Spectrin abundance in TRIM mice at 70 dpt. There was no difference at 140 dpt. ( D , H ) Representative immunoblots of LC3B II and mean densitometric data at ( D ) 70 dpt, revealed that TRIM reduced autophagosome number. No differences were observed at ( H ) 140 dpt. ( n = 7–8/group); summary values are means ± SEM. Comparisons made vs. vehicle controls by two-tailed Student’s t -test; p < 0.05 = significant.

Article Snippet: Primary antibodies of interest were; Alpha II spectrin (RRID: AB_2194351, Cat.# sc-48382; Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:250 incubated overnight at 4 °C then 3 h at room temperature secondary concentration 1:5000) and LC3B (RRID: AB_915950, Cat.# 2775; Cell Signaling Technology, Danvers, MA, USA, secondary concentration 1:5000).

Techniques: Comparison, Saline, Activity Assay, Western Blot, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: Impaired motor activity in a CRISPR SCA5 L253P knock-in mouse is associated with selective β-III-spectrin subcellular redistribution in the cerebellum

doi: 10.64898/2026.03.14.711824

Figure Lengend Snippet: (A) RT-qPCR shows a modest ~20% increase in homozygous β-III-spectrin transcript levels, n=3-4 (B-D) Western blot of cerebellum extracts shows progressive decrease in β-III-spectrin protein in heterozygous and homozygous mice in both soluble and insoluble fractions, n=3. (A, C, D) One way ANOVA comparing to wild-type using Dunnett’s multiple comparison correction.

Article Snippet: Soluble and insoluble samples were resolved by SDS-PAGE gel followed by transferring to Immoblin-FL membranes (Millipore). β-III-spectrin proteins were detected by probing with anti-β-III-spectrin antibody (1:100; Santa Cruz A-8) diluted in 1% casein/1x PBS and 0.1% Tween-20 and IRDye 800CW secondary antibody (1:5000; LICORbio) diluted in 0.1% Tween-20, 0.01% SDS, 1% casein, 1x PBS.

Techniques: Quantitative RT-PCR, Western Blot, Comparison

Journal: bioRxiv

Article Title: Impaired motor activity in a CRISPR SCA5 L253P knock-in mouse is associated with selective β-III-spectrin subcellular redistribution in the cerebellum

doi: 10.64898/2026.03.14.711824

Figure Lengend Snippet: (A) Number of foot slips 6 week and 20 week old mice on 10 or 16 mm, square or round elevated beams. At 6 weeks, no increase in foot slips is observed on any beam. At 20 weeks, β-III-spectrin L253P/L253P mice show increased foot slips on the 10 and 16 mm round and square beams. β-III-spectrin L253P/+ mice show increased number of foot slips that approaches statistical significance on the 10 mm round beam. One way ANOVA followed by multiple comparisons with Tukey’s correction, n=10-14

Article Snippet: Soluble and insoluble samples were resolved by SDS-PAGE gel followed by transferring to Immoblin-FL membranes (Millipore). β-III-spectrin proteins were detected by probing with anti-β-III-spectrin antibody (1:100; Santa Cruz A-8) diluted in 1% casein/1x PBS and 0.1% Tween-20 and IRDye 800CW secondary antibody (1:5000; LICORbio) diluted in 0.1% Tween-20, 0.01% SDS, 1% casein, 1x PBS.

Techniques:

Journal: bioRxiv

Article Title: Impaired motor activity in a CRISPR SCA5 L253P knock-in mouse is associated with selective β-III-spectrin subcellular redistribution in the cerebellum

doi: 10.64898/2026.03.14.711824

Figure Lengend Snippet: (A) Immunofluorescent labelling of β-III-spectrin shows distribution changes in the soma and throughout the dendritic arbor of 20 week mice. (B) Close-up view of soma and proximal dendrites. In WT mice, β-III-spectrin shows a diffuse distribution in the soma and proximal dendrites. In β-III-spectrin L253P/+ mice, β-III-spectrin shows accumulation at the plasma membrane of the soma and proximal dendrites (yellow arrows) and forms small inclusions in the soma (red arrows). In β-III-spectrin L253P/L253P mice, β-III-spectrin localizes strongly to the proximal dendrites and soma plasma membrane with small and large intracellular inclusions present. (C) Quantitation of the total inclusion volume relative to the soma volume in 6 week and 20 week wild-type and homozygous mice. Unpaired t-test, n=3 mice per group with at least 10 somas per image. (D) Hippocampal pyramidal neurons and cortical neurons showing plasma membrane accumulation (yellow arrows) of β-III-spectrin in β-III-spectrin L253P/L253P mice, but no inclusions.

Article Snippet: Soluble and insoluble samples were resolved by SDS-PAGE gel followed by transferring to Immoblin-FL membranes (Millipore). β-III-spectrin proteins were detected by probing with anti-β-III-spectrin antibody (1:100; Santa Cruz A-8) diluted in 1% casein/1x PBS and 0.1% Tween-20 and IRDye 800CW secondary antibody (1:5000; LICORbio) diluted in 0.1% Tween-20, 0.01% SDS, 1% casein, 1x PBS.

Techniques: Clinical Proteomics, Membrane, Quantitation Assay

Journal: bioRxiv

Article Title: Impaired motor activity in a CRISPR SCA5 L253P knock-in mouse is associated with selective β-III-spectrin subcellular redistribution in the cerebellum

doi: 10.64898/2026.03.14.711824

Figure Lengend Snippet: (A) Immunofluorescent labelling of cerebellar sections with nucleoporin 62, β-III-spectrin, and calbindin antibodies shows that inclusions in the β-III-spectrin L253P/L253P tissue localize around the nucleus, in 20 week mice. (B-C) Calbindin labelled Purkinje neurons somas were masked using Imaris software. (B) Labelling with β-III-spectrin antibody and Alexa-488 phalloidin shows F-actin colocalization with the inclusions. (C) Labelling with β-III-spectrin and α-II-spectrin shows α-II-spectrin colocalization with the inclusions.

Article Snippet: Soluble and insoluble samples were resolved by SDS-PAGE gel followed by transferring to Immoblin-FL membranes (Millipore). β-III-spectrin proteins were detected by probing with anti-β-III-spectrin antibody (1:100; Santa Cruz A-8) diluted in 1% casein/1x PBS and 0.1% Tween-20 and IRDye 800CW secondary antibody (1:5000; LICORbio) diluted in 0.1% Tween-20, 0.01% SDS, 1% casein, 1x PBS.

Techniques: Software

Journal: bioRxiv

Article Title: Impaired motor activity in a CRISPR SCA5 L253P knock-in mouse is associated with selective β-III-spectrin subcellular redistribution in the cerebellum

doi: 10.64898/2026.03.14.711824

Figure Lengend Snippet: (A) Immunoprecipitation coupled to tandem mass spectrometry was performed for unbiased identification of proteins that associate with wild-type β-III-spectrin in the cerebellum. STRING identified a cluster of 41 proteins (of 157 total) that are linked to synaptic transmission. Connecting lines represent physical and / or functional linkages between specific proteins, with thicker lines indicating higher confidence interactions. (B) Example proteins from the cluster, with various levels of enrichment, based on normalization abundance values in control versus β-III-spectrin samples. Unpaired t-test, n=3.

Article Snippet: Soluble and insoluble samples were resolved by SDS-PAGE gel followed by transferring to Immoblin-FL membranes (Millipore). β-III-spectrin proteins were detected by probing with anti-β-III-spectrin antibody (1:100; Santa Cruz A-8) diluted in 1% casein/1x PBS and 0.1% Tween-20 and IRDye 800CW secondary antibody (1:5000; LICORbio) diluted in 0.1% Tween-20, 0.01% SDS, 1% casein, 1x PBS.

Techniques: Immunoprecipitation, Mass Spectrometry, Transmission Assay, Functional Assay, Control