Journal:

Article Title: Bcl-2 antiapoptotic protein mediates verotoxin II-induced cell death: possible association between Bcl-2 and tissue failure by E. coli O157:H7

doi:

Figure Lengend Snippet: VTII-Bcl-2 complex formation. (A) VTII, but not VTI, contains the pentameric sequence from the Bcl-2 BH1 domain. Shown are the schematic model, sequence alignment, and protein identities and similarities of human Bcl-2, VTI and VTII. The dotted line indicates the region of splicing variants. (B,C). VTII, but not VTI, interacts with Bcl-2. Expression of Bcl-2 in V (-) and B10 (+) cells was detected with immunoblotting (IB). B10 (B) and/or V (B,C) cell extracts were separated by SDS-PAGE, and far-western blotting (FW) was performed with biotinylated peptides (NWGRL and NWGRI in B) or VTs (VTI and VTII in C). Interacted protein was detected with avidin-HRP and ECL. Bars on the left show positions of protein markers 46, 30, 20, and 14 kDa, from top to bottom. (D) VTII interacts with Bcl-2 through the NWGRI sequence. Far-western blotting analysis was performed with biotinylated VTII and B10 cell extracts in the presence of NWGRI (0, 1, 5, and 10 nm). (E) Coimmunoprecipitation analysis. V (-) or B10 (+) cells were treated with VTII, and then immunoprecipitates with antibodies for Bcl-2 (Bcl-2 IPs) or VTII (VTII IPs) were separated on SDS-PAGE, and immunoblotting analysis with VTII antibody was performed (VTII WB).

Article Snippet: After washing with Tween-PBS, the membranes were incubated with a 1000-fold diluted biotinylated second antibody, washed with Tween-PBS, and then incubated with avidin-HRP (Vector Laboratories, Burlingame, CA) at room temperature for 1 hour.

Techniques: Sequencing, Expressing, Western Blot, SDS Page, Far Western Blot, Avidin-Biotin Assay

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