Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Diet-induced aortic valve disease in mice haploinsufficient for the Notch pathway effector RBPJK/CSL.

doi: 10.1161/ATVBAHA.111.227561

Figure Lengend Snippet: Figure 2. Greater leaflet thickness and increased macrophage deposition in RBPKO/ mice fed an HCVD diet. Shown are Masson Trichrome staining (A to F) of aortic valves indicating collagen deposition (arrows) and Mac3 immuno- staining (G to L) of aortic valves demon- strating macrophage infiltration (red; arrows indicate positive immunostaining) in WT mice fed the CC diet (A and G) or the HCVD diet (D and J), Notch1KO/ mice fed the CC diet (B and H) or the HCVD diet (E and K), and RBPKO/ mice fed the CC diet (C and I) or the HCVD diet (F and L). M and N, Mean data of cusp area (M) and percentage of positively stained sections for Mac3 (N) for all groups (n4 per group). One-way ANOVA and LSD post hoc analysis were used to compare differences between groups. Bars with different letters indi- cate significant differences (P0.05) vs all other groups (LSD post hoc analysis). Scale bar0.1 mm.

Article Snippet: Inflammation was detected with anti-Mac3 rat monoclonal antibody (SC19991, Santa Cruz Technol, 1:200).

Techniques: Staining, Immunostaining

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Methods for investigating STAT3 regulation of lysosomal function in mammary epithelial cells

doi: 10.1007/s10911-024-09563-3

Figure Lengend Snippet: Schematic overview of the lysosomal leakiness assay. The permeability of isolated lysosomes can be assessed by western blotting for cathepsin B and L antibodies to visualise the degree of leakiness of cathepsin hydrolases into the supernatant fraction under different treatment conditions (i.e. vehicle and OSM stimulated conditions in EpH4 cells). Immunoblotting for LAMP2, a lysosomal membrane protein, can be used to standardise the amount of lysosomal material in a pellet during the assay. Alternatively, cathepsin activity levels can be assessed in supernatant and pellet fractions using enzyme activity assays

Article Snippet: Primary and secondary antibodies as required e.g. rat anti-LAMP2 antibody (GL2A7, ab13524, Abcam, 1:200), rat anti-LAMP1 (ID4B, Developmental Studies Hybridoma Bank, 1:200).

Techniques: Permeability, Isolation, Western Blot, Membrane, Activity Assay

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Methods for investigating STAT3 regulation of lysosomal function in mammary epithelial cells

doi: 10.1007/s10911-024-09563-3

Figure Lengend Snippet: STAT3-mediated upregulation of the vesicular compartment in EpH4 cells. A Fluorescence microscopy of LAMP1 and LAMP2 immunostaining in EpH4 cells treated with OSM for 72 h show upregulation of the lysosomal compartment with STAT3 activation. Scale bars: 20 μm. B Fluorescence microscopy of LysoTracker® staining in EpH4 cells treated with OSM for 72 h show upregulation of the acidic compartment with STAT3 activation. Scale bars: 20 μm. C Transmission electron microscopy images of EpH4 cells showing an increased number of degradative vesicles after 72 h of OSM stimulation. Scale bars: 500 nm

Article Snippet: Primary and secondary antibodies as required e.g. rat anti-LAMP2 antibody (GL2A7, ab13524, Abcam, 1:200), rat anti-LAMP1 (ID4B, Developmental Studies Hybridoma Bank, 1:200).

Techniques: Fluorescence, Microscopy, Immunostaining, Activation Assay, Staining, Transmission Assay, Electron Microscopy