Journal: iScience

Article Title: MircroRNA-92b as a negative regulator of the TGF-β signaling by targeting the type I receptor

doi: 10.1016/j.isci.2023.108131

Figure Lengend Snippet:

Article Snippet: Human renal glomerular endothelial cells (HRGECs) (ScienCell, Carlsbad, CA, USA; Cat. #4000) were cultured in Endothelial Cell Medium (ScienCell, Cat.#1001) and human mesangial mells (HMCs) (ScienCell, Cat. #4200) were cultured in Mesangial Cell Medium (ScienCell, Cat. #4201).

Techniques: Recombinant, Control, Plasmid Preparation, Software, Modification, Staining, Extraction, Bicinchoninic Acid Protein Assay, Western Blot, cDNA Synthesis, SYBR Green Assay, Immunohistochemistry, Isolation

Journal: bioRxiv

Article Title: Accumulation of Treg cells is detrimental in late-onset (aged) mouse model of multiple sclerosis

doi: 10.1101/2021.12.16.472986

Figure Lengend Snippet: (A) Workflow of EAE induction, transient inhibition of pTreg cells, blood collection time-points (red arrowheads) for Treg cell tests, and demyelination assay. (B) Effects of transient inhibition of pTreg cells in aged mice. At the first evaluation (one day before the first injection of P300i or vehicle), the percentages of pTreg cells in the two aged groups were higher than the young (left panel). At the second evaluation (one day after the last injection of P300i or vehicle), the percentages of pTreg cells and expression (MFI) of FoxP3 in the aged group treated with P300i (black triangles/line) were significantly reduced. At the third evaluation (12 days after the last treatment), the pTreg cells in the P300i-treated aged group were restored to the same levels as the vehicle-treated control aged group (blue dots/line). All data are expressed as mean ± SEM and were analyzed by One-way ANOVA followed by Dunnett’s multiple post-hoc test. * p < 0.05 young mice +vehicle v.s. aged mice +vehicle; ** p < 0.01 young mice +vehicle v.s. aged mice +vehicle; **** p < 0.0001, young mice +vehicle v.s. aged mice +vehicle; ## p < 0.01, aged mice + vehicle v.s. aged mice + P300i; #### p < 0.0001, aged mice + vehicle v.s. aged mice + P300i. Animal numbers in each group are listed in each panel. (C) Alleviation of the symptoms in late-onset EAE of aged mice. The EAE scores of 5x P300i-treated aged group (black triangles/line) were significantly reduced compared to their counterparts, the vehicle-treated control aged group (blue dots/line). The p -values were calculated using Kruskal-Wallis test followed by Dunnett’s multiple comparisons post hoc test for pairwise comparisons of groups. (D) Illustration of a mouse brain and spinal cord indicating the thoracic segment of the spinal cord for LFB-eosin staining (rightmost illustration). Three representative spinal cord section images of LFB-eosin staining, showing alleviation of demyelination in the spinal cords of aged mice treated with 5x P300i (right image), compared to their counterparts, the vehicle-treated age-matched control mice (middle image). The dotted outlines indicate areas of large foci of demyelination, and blue arrow heads show small foci of demyelination. This experiment was repeated three times with three mice in each group with essentially identical results.

Article Snippet: As shown in , 12 days after EAE induction in young and aged WT mice, P300i (C646; Tocris Bioscience, Cat# 4200) was i.p. injected at 8.9 mg/kg body weight per day for 5 consecutive days (5x) in one of the aged mouse groups similar to published protocol .

Techniques: Inhibition, Injection, Expressing, Control, Staining

Journal: bioRxiv

Article Title: Accumulation of Treg cells is detrimental in late-onset (aged) mouse model of multiple sclerosis

doi: 10.1101/2021.12.16.472986

Figure Lengend Snippet: (A) Flow cytometry gating strategies show the frequencies of peripheral blood Treg cells in the three groups of mice 11-DPI (one day before p300i or vehicle treatment), 17-DPI (one day after the last p300i or vehicle treatment) and 28-DPI (12 days after the last p300i or vehicle treatment). (B) Flow cytometry gating strategies show representative FoxP3+ gates (bottom panels) from CD4 + CD25 + gates of the peripheral blood Treg cells in the three groups of mice 11-DPI, 17-DPI, and 28-DPI.

Article Snippet: As shown in , 12 days after EAE induction in young and aged WT mice, P300i (C646; Tocris Bioscience, Cat# 4200) was i.p. injected at 8.9 mg/kg body weight per day for 5 consecutive days (5x) in one of the aged mouse groups similar to published protocol .

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: Accumulation of Treg cells is detrimental in late-onset (aged) mouse model of multiple sclerosis

doi: 10.1101/2021.12.16.472986

Figure Lengend Snippet: The workflow of the pTreg inhibition is shown in . (A) Expression of IFN-g + in CP-residing CD45 + cells were analyzed one day after the last P300i treatment ( , the second test red arrow). A representative histogram shows IFN-g + CD45 + cells residing at the CP isolated from the brains of young and aged mice treated with P300i or vehicle. Percentages of IFN-g + cells in CD45 + cells and MFI of IFN-g expression are listed in the table (right). (B) Flow cytometry gating strategies of pan-Treg and MOG-specific Treg cells, similar to , in the CNSs of young and aged mice with P300i- or vehicle-treatment. (C) Summarized results of the percentages of pan- (striped bars) and MOGspecific (opened bars) Treg cells (left panel) and Teff cells (middle panel), and ratios of Treg/Teff cells (right panel) in the CNS (a combination of the brain and the spinal cord) among young (cherry) and aged mice, treated with vehicle (blue) or with P300i (black). (D) Flow cytometry gating strategies show representative FoxP3 + gates (bottom panels) from CD4 + CD25 + gates of the CNS (brain and spinal cord) of the three groups of mice. (E) Summarized results of the RQ-MFI of FoxP3 expression in pan- (striped bars) and MOGspecific (open bars) CD4 + CD25 + population in the CNS among young (cherry) and aged mice treated with vehicle (blue) or P300i (black). In panels C and E, each symbol represents an individual animal sample. Data are expressed as mean ± SEM. The p -values between three groups were analyzed by one-way ANOVA with a Dunnett’s multiple post-hoc test, and a statistically significant difference was considered to be p < 0.05, “N.S.” stands for “not significant”.

Article Snippet: As shown in , 12 days after EAE induction in young and aged WT mice, P300i (C646; Tocris Bioscience, Cat# 4200) was i.p. injected at 8.9 mg/kg body weight per day for 5 consecutive days (5x) in one of the aged mouse groups similar to published protocol .

Techniques: Inhibition, Expressing, Isolation, Flow Cytometry