Journal: World Journal of Diabetes

Article Title: Oral Akkermansia muciniphila may ameliorates immune dysregulation in a murine model of streptozotocin-induced type 1 diabetes

doi: 10.4239/wjd.v16.i12.111771

Figure Lengend Snippet: Analysis of the regulatory T cells phenotype, spleen cytokine profile and intestinal mucosa immunohistochemistry (30-fold) in each group of mice. A: Flow gating strategy; B: Columnar diagram of cluster of differentiation (CD) 4 + forkhead box P3 (FoxP3 + ) regulatory T (Treg) cells expression in the spleens of the mice in each group ( n = 8); C: Spleen CD4 + /CD8 + cell ratio histogram for each group ( n = 8); D: Histogram of transforming growth factor-beta (TGF-β) expression in the spleens of the mice in each group ( n = 10); E: Correlation analysis between CD4 + FoxP3 + Tregs and TGF-β ( n = 20); F: Columnar diagram of tumor necrosis factor-alpha expression in the spleens of the mice in each group ( n = 6-7); G: Spleen interferon-gamma expression histogram for each group of mice ( n = 6-7); H: Histogram of interleukin-4 expression in the spleens of the mice in each group ( n = 6-7); I: Nuclear factor kappa-B (NF-κB) p65 positive expression score for the mouse colon (30 times); J: Histogram of NF-κB p65 comprehensive positive intensity histochemistry score for the colon in each group ( n = 4); K: Signal transducer and activator of transcription 1 positive expression score in the mouse colon (30 times); L: Histochemical score of colon signal transducer and activator of transcription. a P < 0.05 vs negative control group. b P < 0.01 vs negative control group. c P < 0.001 vs negative control group. d P < 0.05 vs streptozotocin-induced type 1 diabetes mellitus group. e P < 0.01 vs streptozotocin-induced type 1 diabetes mellitus group. f P < 0.001 vs streptozotocin-induced type 1 diabetes mellitus group. NC group: Negative control group; STZ group: Streptozotocin-induced type 1 diabetes mellitus group; A. muciniphila group: Akkermansia muciniphila intervention group; FSC-A: Forward scatter area; SSC-A: Side scatter area; FSC-H: Forward scatter height; CD: Cluster of differentiation; FoxP3: Forkhead box P3; Treg: Regulatory T; TGF-β: Transforming growth factor-beta; TNF-α: Tumor necrosis factor-alpha; IFN-γ: Interferon-gamma; IL-4: Interleukin-4; NF-κB: Nuclear factor kappa-B; STAT1: Signal transducer and activator of transcription 1.

Article Snippet: The following resources were used: STZ (Sigma, United States), sodium citrate buffer (Shanghai Zeta Company, China), a hematoxylin-eosin staining kit (Wuhan Sevier Biological Company, China), 4% paraformaldehyde fixative (Wuhan Sevier Biological Company, China), an anti-insulin mouse mAb (Wuhan Sevier Biological Company, China), an anti-mouse CD4-fluorescein isothiocyanate antibody (Hangzhou Lianke Biological Company, China), an anti-mouse CD8- APC antibody (Hangzhou Lianke Biological Company, China), an anti-mouse forkhead box P3 (FoxP3)-PE antibody (Hangzhou Lianke Biological Company, China), a purified anti-mouse CD16/32 antibody (Hangzhou Lianke Biological Company, China), a mouse IgG1 isotype control-PE antibody (Hangzhou Lianke Biological Company, China), mouse fluorescence-activated cell sorting concentrate (4 ×) and diluent (Hangzhou Lianke Biological Company, China), rabbit anti-mouse STAT1 polyclonal antibodies (Wuhan Sevier Biotechnology Company, China), rabbit anti-mouse NF-κB p65 polyclonal antibodies (Wuhan Sevier Biotechnology Company, China), a goat anti-mouse immunoglobulin secondary antibody (Wuhan Sevier Biotechnology Company, China), enzyme-linked immunosorbent assay (ELISA) kits (for TNF-α, IFN-γ, IL-4, IL-10, TGF-β, ZO-1 and zonulin; Shanghai Enzymatic Immunobiology Company, China), a DNA extraction kit (Kaijie, Germany), and a double-stranded DNA determination kit (Sigma, United States).

Techniques: Immunohistochemistry, Expressing, Negative Control

Journal: World Journal of Diabetes

Article Title: Oral Akkermansia muciniphila may ameliorates immune dysregulation in a murine model of streptozotocin-induced type 1 diabetes

doi: 10.4239/wjd.v16.i12.111771

Figure Lengend Snippet: Correlation analysis heatmap. a P < 0.05. b P < 0.01. c P < 0.001. CD: Cluster of differentiation; FoxP3: Forkhead box P3; Treg: Regulatory T; TGF-β: Transforming growth factor-beta; TNF-α: Tumor necrosis factor-alpha; IFN-γ: Interferon-gamma; IL-4: Interleukin-4; ZO-1: Zonula occludens-1.

Article Snippet: The following resources were used: STZ (Sigma, United States), sodium citrate buffer (Shanghai Zeta Company, China), a hematoxylin-eosin staining kit (Wuhan Sevier Biological Company, China), 4% paraformaldehyde fixative (Wuhan Sevier Biological Company, China), an anti-insulin mouse mAb (Wuhan Sevier Biological Company, China), an anti-mouse CD4-fluorescein isothiocyanate antibody (Hangzhou Lianke Biological Company, China), an anti-mouse CD8- APC antibody (Hangzhou Lianke Biological Company, China), an anti-mouse forkhead box P3 (FoxP3)-PE antibody (Hangzhou Lianke Biological Company, China), a purified anti-mouse CD16/32 antibody (Hangzhou Lianke Biological Company, China), a mouse IgG1 isotype control-PE antibody (Hangzhou Lianke Biological Company, China), mouse fluorescence-activated cell sorting concentrate (4 ×) and diluent (Hangzhou Lianke Biological Company, China), rabbit anti-mouse STAT1 polyclonal antibodies (Wuhan Sevier Biotechnology Company, China), rabbit anti-mouse NF-κB p65 polyclonal antibodies (Wuhan Sevier Biotechnology Company, China), a goat anti-mouse immunoglobulin secondary antibody (Wuhan Sevier Biotechnology Company, China), enzyme-linked immunosorbent assay (ELISA) kits (for TNF-α, IFN-γ, IL-4, IL-10, TGF-β, ZO-1 and zonulin; Shanghai Enzymatic Immunobiology Company, China), a DNA extraction kit (Kaijie, Germany), and a double-stranded DNA determination kit (Sigma, United States).

Techniques:

Journal: Communications Biology

Article Title: Rectified activity-dependent population plasticity implicates cortical adaptation for memory and cognitive functions

doi: 10.1038/s42003-024-07186-2

Figure Lengend Snippet: A Illustration of the rectified activity-dependent population plasticity (RAPP) and the empirical parameters describe it (Amp & s.d.). S, population response state; Amp., amplitude, the mean response of the subgroup; s.d., standard deviation of the subgroup. B , C Simulation of population dynamics in neocortex. A Gaussian-distributed pattern was seeded in the first step. Activity changes from one step to the next were determined by the empirical parameters. ( B ) Pattern decay. Rows indicate the same neuron; columns indicate sequential steps. The left activity map was plotted by the deviation of each neuron (smallest deviation at top); the right activity map was sorted according to the signal intensity in the first step. Representative patterns in steps were enlarged in separated columns. Scatter plots show the decay of patterns by loss of correlation over increasing time steps. C Dynamics of individual neurons in adaptation. The right activity map was sorted according to the position of the peak activity step for each neuron. Small panels show the enlarged sector indicated as the gray and black bars. D – F Dynamics of cortical memory traces. D Mice were subjected to several trials of contextual fear conditioning training or memory recall over one month. Representative images of EGR1-EGFP signal in recent and early recall trials in visual cortex. Red, EGFP signal in homecage trial of day 0; Green, EGFP signal in context A recall trials. Arrowheads indicate the trace neuron of day 4 and arrows indicate the trace neurons of day 17. Scale bar, 40 μm. E , F Simulated activities of individual neurons (for details see “Methods” section) ( E ) and recorded memory trace responses over 30 days ( F ). Neurons counted as traces are the most active neurons on the referenced day (Δ F over 3x of standard deviation). In the simulation ( E ), neurons counted as traces are the most active neurons on the referenced cycle (Δ F over 3 x of standard deviation, on 20th, 40th, and 140th round of updating). Simulated activities in those three cell populations are shown. Early phase, day 2–4 traces vs. later phase, day 7–28 traces: n > 80 memory trace neurons from 6 mice for each group; *** p < 0.001, two-way ANOVA. Error bars, s.e.m.

Article Snippet: Training consisted of a 3 min exposure of animals to the conditioning box (Multi-conditioning system, TSE system) followed by a foot shock (2 sec, 0.8 mA, constant current).

Techniques: Activity Assay, Standard Deviation

Journal: Communications Biology

Article Title: Rectified activity-dependent population plasticity implicates cortical adaptation for memory and cognitive functions

doi: 10.1038/s42003-024-07186-2

Figure Lengend Snippet: A , B Simulation of the pattern similarity decay using the empirical parameters of the population plasticity. A Gaussian-distributed activity pattern was used as the initial seed. The pattern in the third cycle was treated as the initial representation. α, is the slope parameter. C Representative example of pattern similarity decay after repeated CtxA exposure in VIS cortex of mice. The mouse explored the box A for 3 min each day. Data was fitted with an exponential curve (red). D Freezing in box A after contextual fear conditioning in hippocampus-lesion group ( n = 5 mice) and normal group ( n = 9 mice), Two-way ANOVA test, group p < 0.0001. E Representative example of population activity pattern decay in the VISam of normal mice (Blue) and HC-lesion mice (Red). F Quantifications of the half-life for the pattern decay in each recorded areas of normal mice (Control) and HC-lesion mice. Context only group, n = 6 mice; contextual fear group, normal, n = 9 mice; HC-lesion, n = 5 mice. Multiple cortices were recorded from one mouse. The roof of half-life was set to 40 days. Error bars, s.e.m.

Article Snippet: Training consisted of a 3 min exposure of animals to the conditioning box (Multi-conditioning system, TSE system) followed by a foot shock (2 sec, 0.8 mA, constant current).

Techniques: Activity Assay, Control

Journal: bioRxiv

Article Title: Cellular toxicity of iHAP1 and DT-061 does not occur through PP2A-B56 targeting

doi: 10.1101/2021.07.08.451586

Figure Lengend Snippet: A) Analysis of the purity of PP2A holoenzyme preparations by colloidal staining and Western blotting. Methylation of the C-tail of PPP2C was confirmed using an antibody against CH 3 - L309. Samples were analysed by mass spectrometry which confirmed that no other protein phosphatases were present in the preparations. B) Titration experiment of different PP2A holoenzyme preparations to determine appropriate concentration to use in enzymatic assays. C-D) In vitro dephosphorylation assays using model phosphopeptides and purified PP2A holoenzymes. The phosphopeptides were incubated with specific PP2A holoenzymes together with iHAP1 or DT-061 as indicated. The reaction was stopped after 15 minutes, and the amount of phosphate released was measured. Mean and SD from three technical replicates of one out of three independent experiments is shown. E) In vitro dephosphorylation assays using a different preparation of PP2A holoenzymes as well as a PP2A-B56α complex reconstituted by expressing all 3 subunits in HEK-293T cells.

Article Snippet: Filming was performed on a LSM 880 Airyscan Confocal attached to an inverted stand Zeiss AxioObserver.Z1 (Carl Zeiss), equipped with an Incubation box XL Multi S1 set to 37°C, and sample was mounted on a C-Apochromat x40/1.2W objective.

Techniques: Staining, Western Blot, Methylation, Mass Spectrometry, Titration, Concentration Assay, In Vitro, De-Phosphorylation Assay, Purification, Incubation, Expressing