Journal: Scientific reports

Article Title: Mutant and curli-producing E. coli enhance the disease phenotype in a hSOD1-G93A mouse model of ALS.

doi: 10.1038/s41598-023-32594-5

Figure Lengend Snippet: Figure 3. Gait kinematics and locomotion characteristics. (A–D) Average running speed and stride time measured with TreadScan analyses showed significant slowing of pace in hSOD1 males compared to WT controls (A). This finding was absent in females (C). Stride time, defined as the time elapsed between two successive initiations of stances, showed significant increases in hSOD1 males compared to WT controls (B). This finding was absent in females (D). Data represented as mean + /-SEM for (A–D). (E) Within the male hSOD1 cohort, the ANOVA was significant for effect of time on running speed, but bacterial feeding did not have a significant effect. (F) Bacterial feeding also did not significantly effect stride time. Within the male hSOD1 group, curli-fed mice showed significant differences for maximum lateral deviation of hind feet from the axis of the body (G), inability to efficiently move body from a straight axis (H), and a smaller print area of the hind feet (I) towards later months. *p < 0.05, Repeated measures two-way ANOVA or mixed-effects model analyses (REML) with Tukey’s multiple comparisons. n = 6–8 per group.

Article Snippet: Tissue sections were deparaffinized, rehydrated and probed with the following antibodies: hSOD1 (SOD1 MS785, GTX57211), ChAT (ProteinTech, 20,747-1-AP), GFAP (ABclonal, A10873), Iba1 (Abcam, ab17886), Histochemistry was performed using antigen retrieval with citric acid buffer at 95C for 30 min and Vector ABC and NovaRed substrate system according to manufacturer instructions (Vector Laboratories).

Techniques:

Journal: Scientific reports

Article Title: Mutant and curli-producing E. coli enhance the disease phenotype in a hSOD1-G93A mouse model of ALS.

doi: 10.1038/s41598-023-32594-5

Figure Lengend Snippet: Figure 5. Neurodegeneration, inflammation, and demyelination in the nervous system. (A,B) Within the male WT group, mutant and curli-fed mice showed significantly decreased cholinergic (ChAT+) neurons compared to vehicle. Within the vehicle group, hSOD1 males had fewer ChAT+ neurons compared to WT controls. (C,D) Within the male hSOD1 cohort, curli-fed mice showed increased astrogliosis in brainstem compared to vehicle. Images were quantified in ImageJ utilizing percent positive signal of staining. (E,F) Within the male hSOD1 cohort, mutant and curli-fed mice showed significant demyelination in white matter of spinal cords compared to vehicle groups (encroachment of pink cytoplasmic stain eosin into areas of Luxol Fast Blue stained myelin). *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. n = 3–5 per group.

Article Snippet: Tissue sections were deparaffinized, rehydrated and probed with the following antibodies: hSOD1 (SOD1 MS785, GTX57211), ChAT (ProteinTech, 20,747-1-AP), GFAP (ABclonal, A10873), Iba1 (Abcam, ab17886), Histochemistry was performed using antigen retrieval with citric acid buffer at 95C for 30 min and Vector ABC and NovaRed substrate system according to manufacturer instructions (Vector Laboratories).

Techniques: Mutagenesis, Staining

Journal: Journal of pharmacological sciences

Article Title: Schwann cell derived-peroxiredoxin protects motor neurons against hydrogen peroxide-induced cell death in mouse motor neuron cell line NSC-34.

doi: 10.1016/j.jphs.2023.07.006

Figure Lengend Snippet: Fig. 4. Expression of PRDX isoforms in the lumbar spinal cord of WT and superoxide dismutase 1 (SOD1)G93A transgenic (Tg) mice. a: Photographs of 1e3, 4e6, and 7e9 panels show double immunostaining for PRDX with glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), and NeuN, respectively, in the lumbar spinal cord of WT mice at 10 weeks of age. Photographs of panels 10 and 11 show immunostaining in continuous sections with PRDX or ionized calcium binding adaptor molecule (Iba1) in the lumbar spinal cord of WT mice at 10 weeks old. The boundary between the white and grey matter of the lumbar spinal cord is indicated by a dotted line. Representative immunofluorescence images display PRDX (red; panels 2, 5, and 8) with GFAP (green; panel 1), MBP (green; panel 4), and NeuN (green; panel 7). The merge diagram in the rightmost panels is a 2-overlay of the diagram

Article Snippet: Human WT SOD1 pCMV6-XL5 plasmid and recombinant mouse PRDX1 protein (AR52067PU-N) were purchased from ORIGENE (Rockville, MD, USA).

Techniques: Expressing, Transgenic Assay, Double Immunostaining, Immunostaining, Binding Assay

Journal: Journal of pharmacological sciences

Article Title: Schwann cell derived-peroxiredoxin protects motor neurons against hydrogen peroxide-induced cell death in mouse motor neuron cell line NSC-34.

doi: 10.1016/j.jphs.2023.07.006

Figure Lengend Snippet: Fig. 5. The role of WT and SOD1G93A transduction in YST-1 cells on the expression of PRDX isoforms in YST-1 cells and the protective effect of YST-1 CM against hydrogen peroxide- induced cell death in NSC-34 cells. In all figures, the data represent means ± SEM. a: YST-1 cells were transfected with vectors containing human WT SOD1 or SOD1G93A. Forty-eight hours after transfection, SOD1 protein expression was validated by Western blot analysis (n ¼ 3). b-actin was used as a loading control. Relative SOD1 protein levels were quantified, and the graphs show the correct bands. Mock: a band of SOD1 from empty vector-transfected YST-1 cells. Data are expressed as arbitrary units and represent the mean ± SEM *P < 0.05, significantly different from empty vector-transfected YST-1 cells; bee: Forty-eight hours after transfection, the mRNA expression of (b) PRDX1, (c) PRDX2, (d) PRDX3, and (e) PRDX4 was examined by real-time RT-PCR (n ¼ 4). Data are expressed as arbitrary units normalised to b-actin and are represented as mean ± SEM. *P < 0.05, significantly different from WT SOD1-transfected YST-1 cells; f: Forty-eight hours after transfection, PRDX1e4 protein expression was validated by Western blot analysis (n ¼ 4). b-actin was used as a loading control. Relative PRDX1e4 protein levels were quantified, and their graphs are shown below the bands. The control showed the PRDX1e4 band in YST-1 cells without transfection. Data are expressed as arbitrary units and represent the mean ± SEM. *P < 0.05, significantly different from WT SOD1-transfected YST-1 cells; g: NSC-34 cells were treated with hydrogen peroxide (100 mM) for 24 h in the absence or presence of CM prepared from YST-1 cells transfected with the WT and SOD1G93A expression vectors. *P < 0.05 and **P < 0.01, significant difference between bracketed values.

Article Snippet: Human WT SOD1 pCMV6-XL5 plasmid and recombinant mouse PRDX1 protein (AR52067PU-N) were purchased from ORIGENE (Rockville, MD, USA).

Techniques: Transduction, Expressing, Transfection, Western Blot, Control, Plasmid Preparation, Quantitative RT-PCR

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: Immunodepletion of SOD1 seeding activity using anti-SOD1 antibodies. a SOD1 RT-QuIC of supernatants of sALS spinal cord homogenate exposed to beads alone (green), or beads linked to either a pan-SOD1 antibody (pink) or an isotype-control antibody (gray). b Same as panel a , except for the use of the C4F6 SOD1 antibody (orange) and a different isotype-control antibody treatment (gray). A non-ALS negative control spinal cord homogenate was included without antibody treatment and remained ThT-negative throughout. All samples at 5 × 10 –3 spinal cord tissue dilutions. ThT fluorescence traces from individual quadruplicate wells are shown

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Immunodepletion, Activity Assay, Control, Negative Control, Fluorescence

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: SOD1 RT-QuIC parameters for immunodepletion of sALS spinal cord homogenate

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Immunodepletion

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: SOD1 RT-QuIC plot of cervical spinal cord homogenates from SOD1 familial ALS patients 2 ( a ) and 5 ( b ) at 10 –3 —10 –5 dilutions. Data from non-ALS control cervical spinal cord (10 –3 dilution) shown

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Control

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: SOD1 RT-QuIC plot of sporadic ALS cases using cervical and thoracic spinal cord homogenates. In a , sporadic ALS patients 5 (cervical) and 6 (thoracic) are shown, and in b , sporadic ALS patients 3 (cervical) and 8 (thoracic) are shown at the designated dilutions. Data from non-ALS control spinal cords are shown for comparison

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Control, Comparison

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: SOD1 RT-QuIC plot of cervical spinal cord homogenates from C9ORF72 fALS patients 2 ( a ) and 4 ( b ) compared to non-ALS control spinal cord at the designated dilutions

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Control

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: Receiver operating characteristic curves for SOD1 RT-QuIC assay at 10 –3 spinal cord dilutions (threshold 5000 RFU and 125 h) ( a ) and 10 –4 spinal cord dilutions (threshold 5000 RFU and 175 h) ( b )

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques:

Journal: Acta Neuropathologica

Article Title: Seeding activity of human superoxide dismutase 1 aggregates in familial and sporadic amyotrophic lateral sclerosis postmortem neural tissues by real-time quaking-induced conversion

doi: 10.1007/s00401-024-02752-8

Figure Lengend Snippet: SOD1 RT-QuIC parameters (mean ± standard deviation) on patient neural tissue

Article Snippet: Preparation of human SOD1 plasmid cDNA encoding full length WT human SOD1 was manufactured by Genscript (Piscataway NJ, USA) and placed in the pET 28a(+) vector with an N-terminal 6 × His-tag.

Techniques: Standard Deviation, Fluorescence, Negative Control

Journal: Scientific reports

Article Title: Mutant and curli-producing E. coli enhance the disease phenotype in a hSOD1-G93A mouse model of ALS.

doi: 10.1038/s41598-023-32594-5

Figure Lengend Snippet: Figure 3. Gait kinematics and locomotion characteristics. (A–D) Average running speed and stride time measured with TreadScan analyses showed significant slowing of pace in hSOD1 males compared to WT controls (A). This finding was absent in females (C). Stride time, defined as the time elapsed between two successive initiations of stances, showed significant increases in hSOD1 males compared to WT controls (B). This finding was absent in females (D). Data represented as mean + /-SEM for (A–D). (E) Within the male hSOD1 cohort, the ANOVA was significant for effect of time on running speed, but bacterial feeding did not have a significant effect. (F) Bacterial feeding also did not significantly effect stride time. Within the male hSOD1 group, curli-fed mice showed significant differences for maximum lateral deviation of hind feet from the axis of the body (G), inability to efficiently move body from a straight axis (H), and a smaller print area of the hind feet (I) towards later months. *p < 0.05, Repeated measures two-way ANOVA or mixed-effects model analyses (REML) with Tukey’s multiple comparisons. n = 6–8 per group.

Article Snippet: Tissue sections were deparaffinized, rehydrated and probed with the following antibodies: hSOD1 (SOD1 MS785, GTX57211), ChAT (ProteinTech, 20,747-1-AP), GFAP (ABclonal, A10873), Iba1 (Abcam, ab17886), Histochemistry was performed using antigen retrieval with citric acid buffer at 95C for 30 min and Vector ABC and NovaRed substrate system according to manufacturer instructions (Vector Laboratories).

Techniques:

Journal: Scientific reports

Article Title: Mutant and curli-producing E. coli enhance the disease phenotype in a hSOD1-G93A mouse model of ALS.

doi: 10.1038/s41598-023-32594-5

Figure Lengend Snippet: Figure 5. Neurodegeneration, inflammation, and demyelination in the nervous system. (A,B) Within the male WT group, mutant and curli-fed mice showed significantly decreased cholinergic (ChAT+) neurons compared to vehicle. Within the vehicle group, hSOD1 males had fewer ChAT+ neurons compared to WT controls. (C,D) Within the male hSOD1 cohort, curli-fed mice showed increased astrogliosis in brainstem compared to vehicle. Images were quantified in ImageJ utilizing percent positive signal of staining. (E,F) Within the male hSOD1 cohort, mutant and curli-fed mice showed significant demyelination in white matter of spinal cords compared to vehicle groups (encroachment of pink cytoplasmic stain eosin into areas of Luxol Fast Blue stained myelin). *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. n = 3–5 per group.

Article Snippet: Tissue sections were deparaffinized, rehydrated and probed with the following antibodies: hSOD1 (SOD1 MS785, GTX57211), ChAT (ProteinTech, 20,747-1-AP), GFAP (ABclonal, A10873), Iba1 (Abcam, ab17886), Histochemistry was performed using antigen retrieval with citric acid buffer at 95C for 30 min and Vector ABC and NovaRed substrate system according to manufacturer instructions (Vector Laboratories).

Techniques: Mutagenesis, Staining