Journal: Dermatology and Therapy

Article Title: A Nonsteroidal Novel Formulation Targeting Inflammatory and Pruritus-Related Mediators Modulates Experimental Allergic Contact Dermatitis

doi: 10.1007/s13555-018-0223-8

Figure Lengend Snippet: Naïve control ears exhibit dense sensory fibers and a distinctive PAR-2 band. a Merged immunohistochemistry image showing sensory fibers (green) within the epidermis and distinctive PAR-2 labeling (red). b Green fluorescence channel revealing PGP 9.5 plus beta III tubulin-labeled complex sensory fibers within the epidermis, arising from underlying fiber tracts within the dermis. c Red fluorescence channel demonstrating a narrow bright dense PAR-2-labeled band near the epidermis/dermis interface. Light background label also appears within the stratum corneum of all immunopreparations. d Blue fluorescence channel showing the DAPI-labeled nuclei within this section. Scale bar, 50 μm. e Dermatitic ears generally exhibit increases in PAR-2 deeper within the tissue, when compared to naïve control and vehicle-alone ears (red label), indicating that inflammation has been triggered. The green channel reveals the extent of the sensory fibers within the epidermis, also seen within the merged images at the right. Scale bar, 25 μm

Article Snippet: Following Triton X-100 permeablization and blocking, samples were incubated with PGP 9.5 (1:500, Abcam, ab108986), beta III tubulin (1:200, Abcam, ab52901), and PAR-2 (1:200, Santa Cruz) in blocking buffer for 3 days, washed, and then incubated for 1 h in donkey anti-rabbit IgG Alexa Fluor 488 (Invitrogen) for simultaneous labeling of both PGP 9.5 and beta III tubulin, and donkey anti-goat IgG Alexa Fluor 633 (Invitrogen) for PAR-2.

Techniques: Immunohistochemistry, Labeling, Fluorescence

Journal: The Journal of Physiology

Article Title: A novel enteric neuron–glia coculture system reveals the role of glia in neuronal development

doi: 10.1113/JP271989

Figure Lengend Snippet: Neuron‐enriched primary culture from E15 embryonic rat intestine A , schematic summarizing the principle of the enteric neuron/glia coculture model. The two cell types are first prepared separately, and are then combined once the neurons have attached to the coverslips. The neurons and the glia remain separated by a narrow gap provided by the paraffin dots stuck on the glass coverslip. B , immunostaining for enteric neurons (Hu) and myofibroblasts (α‐SMA) at 1, 4, 8 and 12 DIV in E15 embryonic rat intestine cultures grown on glass coverslips. Cell nuclei are labelled with DAPI. Scale bar = 50 μm. C , quantification of the percentage (mean ± SEM, n = 6) of enteric neurons, EGCs and myofibroblasts in cultures grown on glass coverslips. D , Western blot with PGP9.5 and GFAP antibodies of lysates from cultures grown on glass coverslips or from mixed cultures. E , neuronal cell death assessed by active caspase 3 immunostaining during in vitro ]

Article Snippet: Membranes were blocked for 1 h at 25°C in Tris‐buffered saline‐Tween 0.1% (TBST) (150 m m NaCl, 15 m m Tris, 4.6 m m Tris base, Tween 0.1%, pH 7.4) containing 5% non‐fat dry milk and incubated overnight at 4°C with the primary antibodies: rabbit anti‐PGP9.5 (Cedarlane, Burlington, Ontario, Canada; diluted 1:5000), rabbit anti‐GFAP (Dako, diluted 1:5000).

Techniques: Immunostaining, Western Blot, In Vitro