Journal: Oxidative Medicine and Cellular Longevity

Article Title: Streptococcus lutetiensis Induces Autophagy via Oxidative Stress in Bovine Mammary Epithelial Cells

doi: 10.1155/2022/2549772

Figure Lengend Snippet: Antibody details.

Article Snippet: Anti-LAMP2 , AF1036 , Beyotime , Shanghai, China , 1 : 1000.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Streptococcus lutetiensis Induces Autophagy via Oxidative Stress in Bovine Mammary Epithelial Cells

doi: 10.1155/2022/2549772

Figure Lengend Snippet: S. lutetiensis decreased pH in lysosomes to further block the autophagy flux. (a) To assess lysosomal pH, cells were stained with 100 nM LysoTracker Deep Red at 37°C for 30 min after being infected with S. lutetiensis . Scale bars: 20 μ m. (b) After being infected with S. lutetiensis , cells were stained with AO at 37°C to assess lysosomal pH. Scale bars: 20 μ m. (c–e) Protein levels of LAMP2, cathepsin D, and cathepsin L in MAC-T cells at various intervals after infection with S. lutetiensis . Upper panels: representative Western blot images; lower panels: quantitative analysis (mean ± SD, n = 3, ∗ represents the significance with the 0 h group, ∗ p < 0.05).

Article Snippet: Anti-LAMP2 , AF1036 , Beyotime , Shanghai, China , 1 : 1000.

Techniques: Blocking Assay, Staining, Infection, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Streptococcus lutetiensis Induces Autophagy via Oxidative Stress in Bovine Mammary Epithelial Cells

doi: 10.1155/2022/2549772

Figure Lengend Snippet: NAC efficiently decreased S. lutetiensis -induced autophagy in MAC-T. (a–f) Protein levels of LC3II/I, p62, Beclin1, LAMP2, CTSD, and CTSL. Upper panels: representative Western blot images; lower panels: quantitative analysis (mean ± SD, n = 3, ∗ represents the significance with the control group, # represents the significance with the treated group, ∗ p < 0.05, # p < 0.05; C: control; T: treatment; R: rapamycin; M: 3-MA; T+N: treatment + NAC). (g) Lysosome detection comparison between infected with S. lutetiensis with/without NAC. Scale bars: 20 μ m.

Article Snippet: Anti-LAMP2 , AF1036 , Beyotime , Shanghai, China , 1 : 1000.

Techniques: Western Blot, Infection

Journal: bioRxiv

Article Title: The non-structural proteins NS3 and NS5A of Hepatitis C Virus (HCV) are degraded by two host proteolysis systems

doi: 10.1101/2023.08.16.553615

Figure Lengend Snippet: A. CLSM analysis of Huh7.5-Jc1 cells after treatment with different autophagy modulators and with DMSO, respectively. NS5A was stained with specific Alexa-633 (cyan blue; top left panel), NS3 with specific Alexa-488 (green; top left panel) and LAMP2 with cy3 (red; middle left panel). The bottom left panel shows an overlay of the two panels and a DAPI DNA-stain (blue). The right panel shows an enlargement of the area indicated in the overlay. Laser intensity and laser gain were kept constant. Magnification of 100X is shown. B. Quantification of the pixel co-localization of NS5A and NS3 with LAMP2 using a tMOC analysis. tMOC values were calculated from 6-8 cells of each replicate (n=3, mean ± standard error). The statistical significance was analyzed with a two-tailed unpaired t -test. * = p<0.05, *** = p<0.001, ns = non-significant.

Article Snippet: Anti-NS3 (Mouse-monoclonal, Abcam, Cambridge, UK), anti-NS5A [ ]; Rabbit-polyclonal), anti-PSMB5 (Rabbit-polyclonal, Abcam, Cambridge, UK, or Mouse-polyclonal, Sigma-Aldrich, Seelze, DE) and anti-LAMP2 (Guinea pig-polyclonal, Progen Biotechnik, Heidelberg, DE) were used as primary antibodies.

Techniques: Staining, Two Tailed Test