Journal: Physiological Reports

Article Title: IMT504 protects beta cells against apoptosis and maintains beta cell identity, without modifying proliferation

doi: 10.14814/phy2.15790

Figure Lengend Snippet: Confocal microscopic analysis of two examples of IMT504‐Texas Red incorporation into MIN6B1 cells after 4 h incubation; nuclei stained with DAPI (blue), 60X, bar = 20 μM.

Article Snippet: Confocal microscopy was performed to localize cells staining for IMT504 (red) and DAPI (nuclear staining, blue, VECTOR laboratories).

Techniques: Incubation, Staining

Journal: Journal of Applied Microbiology

Article Title: Bacillus anthracis spores germinate extracellularly at air–liquid interface in an in vitro lung model under serum‐free conditions

doi: 10.1111/jam.12872

Figure Lengend Snippet: Immunofluorescence of transwell inserts of differentiated normal human bronchial epithelial cells after infection suggest minimal uptake as assessed by localization (yellow) with various cellular markers. DAPI nuclear stain‐ blue; B. anthracis ‐ red; and cell markers (a) green‐ β‐actin (b) green‐β‐tubulin (c) green‐MUC5AC (mucus) all three imaged at 4 h postinfection. (d) 24 h infections, noticeable vegetative growth was evident for BA, green‐ β‐actin for specified image. Images were acquired at either 40× or 67× objectives, Scale bar = 50 microns.

Article Snippet: Secondary antibodies for fluorescent staining were Alexa Fluor ® 594 Goat Anti‐Rabbit IgG (A‐1101; Life Technologies) and Alexa Fluor ® 488 Goat Anti‐Mouse (A‐10680; Life Technologies) both at 2 μ g ml −1 that was subsequently followed with 1 μ g ml −1 DAPI nuclear blue stain (D1306; Life Technologies).

Techniques: Immunofluorescence, Infection, Staining

Journal: Journal of Applied Microbiology

Article Title: Bacillus anthracis spores germinate extracellularly at air–liquid interface in an in vitro lung model under serum‐free conditions

doi: 10.1111/jam.12872

Figure Lengend Snippet: A549 vs normal human bronchial epithelial (NHBE) cells in differentiated ALI state induce faster BA germination and proliferation. (a) Immunofluorescence of transwells at 1–5 h assessed for spore out‐growth indicative of germination. Shown is representative 2 hpi time points. DAPI nuclear stain‐ blue; B. anthracis ‐ red. The ‘halo’ like structure (arrow) commonly seen for nongerminated spores by bright field was also more apparent for NHBE than A549 at early time points and that is reflected at 2 hpi by immunofluorescence. A minimum of 200 bacteria for each time point was counted and the percent of bacillus‐shaped bacteria is shown (right). Spores in A549 cells (grey circles‐ ) and spores in NHBE cells (black triangles‐▲) (b) CFU plates counts from transwells at ALI for differentiated NHBE (black bar‐■) vs NHBE cells in traditional submerged 2D state (white bar‐□) compared to A549 cell infections in differentiated state (horizontal striped bar‐ ) vs A549 undifferentiated state (diagonal striped bar‐ ). Differentiated A549 transwells leaked at 12 h and are therefore labelled as not applicable (NA) due to leaking of basal media at ALI into spore environment. All multiplicity of infections (MOI) were performed at 1·0 with 100 000 spore input.

Article Snippet: Secondary antibodies for fluorescent staining were Alexa Fluor ® 594 Goat Anti‐Rabbit IgG (A‐1101; Life Technologies) and Alexa Fluor ® 488 Goat Anti‐Mouse (A‐10680; Life Technologies) both at 2 μ g ml −1 that was subsequently followed with 1 μ g ml −1 DAPI nuclear blue stain (D1306; Life Technologies).

Techniques: Immunofluorescence, Staining