Journal: bioRxiv

Article Title: Satellite stem cell dysfunction in Duchenne muscular dystrophy

doi: 10.1101/2024.07.24.604963

Figure Lengend Snippet: DMD satellite cells have impaired autophagy and senescence. A) Feature plot showing cells positive for the senescence GO term, showing higher amounts in the regions where cycling progenitors and DMD enriched cells cluster. B-D) ddPCR quantification of gene expression in satellite cells in B) senescence-associated genes Cdkn1a and Cdkn2a from non-injured (NI), one day post injury (1DPI) and three days post injury (3DPI) B10 and mdx mice showing dysregulation in mdx . (F = 101.1, P < 0.0001; F = 89.58, P < 0.0001) C) Strain-comparison of the expression of autophagy-associated genes showing reduced expression of autophagy genes in both DMD models compared to healthy controls (t = 5.260, 1.917; 8.707, 1.513; 3.603, 3.130) and D) changes in autophagy-associated genes in NI, 1DPI and 3DPI B10 and mdx satellite cells demonstrating impaired autophagy dynamics in mdx cells during regeneration ([F,P] = [15.51, 0.0004], [62.45, 0.0001], [6.518, 0.0109], [24.28, <0.0001], [20.22, 0.0001]. E) Representative images of IF staining against PAX7, MYOG and MyHC from mdx primary myoblasts treated with an autophagy inhibitor (3-MA, 5 mM and H 2 O control) or autophagy inducer (Tat-D11, 10 µM and Scramble control) for two hours prior to differentiation for two days. F) Fusion index of each condition was determined and shows increased differentiation after Tat-D11 treatment (t = 0.001 (3MA), 2.894 (Tat-D11)). Scalebars = 25 µM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (two-tailed unpaired t-test ( C, F ) and two-way analysis of variance ( B, D )). Data are expressed as mean ± SD.

Article Snippet: When cells reached 80-90% confluence, cells were treated for two hours with either 10 µM Tat-Beclin 1 D11 peptide (Novus Biologicals) or Tat-Beclin 1 L11S peptide scramble control (vehicle, Novus Biologicals); 5 mM 3-methyladenine (3-MA, Sigma) or water (vehicle) , .

Techniques: Expressing, Comparison, Staining, Control, Two Tailed Test

Journal: Immunity

Article Title: Autophagy modulates endothelial junctions to restrain neutrophil diapedesis during inflammation

doi: 10.1016/j.immuni.2021.07.012

Figure Lengend Snippet: Modulation of EC autophagy controls neutrophil extravasation and cellular tissue damage (A and B) Neutrophil extravasation in chimeric Atg5 fl/fl and Atg5 ΔEC mice (A) treated i.s. with PBS or LPS and (B) subjected to local IR injury (n = 3–8 mice/group). (C) Representative (n = 8) confocal images of IR-stimulated cremasteric PCVs (PECAM-1) immunostained for neutrophils (MRP14) (scale bar, 30 μm). (D) Neutrophil extravasation in Atg5 fl/fl and Atg5 iΔEC mice subjected to local IR injury (n = 6 mice/group). (E) Propidium iodide (PI) + cells in Atg5 fl/fl and Atg5 ΔEC mice subjected to IR injury, as quantified by confocal IVM (n = 3–5 mice/group). (F and G) GFP-LC3 puncta or endogenous LC3 puncta per venular EC area in cremasteric PCVs of (F) GFP-Map1lc3 TG/+ mice and (G) Atg5 fl/fl and Atg5 ΔEC mice treated i.s. with scrambled or Tat-Beclin 1 peptide (n = 3–5 mice/group). (H and I) Neutrophil extravasation in (H) WT mice and (I) Atg5 fl/fl and Atg5 iΔEC mice subjected to local IR injury and treated i.s. with scrambled or Tat-Beclin 1 (n = 4–6 mice/group). (J and K) Intravascular neutrophils in WT mice subjected to local IR injury and treated i.s. with scrambled or Tat-Beclin 1 (n = 3–4 mice/group) (J), and (K) representative confocal images (n = 3–6) of cremasteric PCVs (PECAM-1) immunostained for neutrophils (MRP14) (scale bar, 30 μm). Means ± SEMs. Statistically significant difference from controls or between indicated groups is shown by ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001; ns, not significant. See also Figure S2 .

Article Snippet: Tat-Beclin 1 L11S Peptide - Scrambled Control , Bio-techne , Cat#NBP2-49887.

Techniques:

Journal: Immunity

Article Title: Autophagy modulates endothelial junctions to restrain neutrophil diapedesis during inflammation

doi: 10.1016/j.immuni.2021.07.012

Figure Lengend Snippet:

Article Snippet: Tat-Beclin 1 L11S Peptide - Scrambled Control , Bio-techne , Cat#NBP2-49887.

Techniques: Blocking Assay, Generated, Recombinant, Antibody Labeling, SYBR Green Assay, Luciferase, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software