Journal: PLoS Genetics

Article Title: Pregnancy-Induced Noncoding RNA ( PINC ) Associates with Polycomb Repressive Complex 2 and Regulates Mammary Epithelial Differentiation

doi: 10.1371/journal.pgen.1002840

Figure Lengend Snippet: (A–C) RNA was isolated from FACs sorted mammary populations including, virgin luminal (VL) and basal (VB) as well as pregnant luminal (PL) and basal (PB). (A) qPCR showed Krt8 expression was enriched in the luminal populations (VL and PL), (B) Krt14 was enriched in the basal populations (VB and PB) and (C) mPINC was enriched in the pregnant luminal population, (D–F) MECs were FACs sorted into luminal and basal populations using CD24 and CD29. The luminal population was selected and further sorted into mature luminal (ML), luminal progenitors (LP), and alveolar progenitors (AP) using CD14 and ckit. (D) FACs dot plots showing CD24 and CD29 (left panel) as well as CD14 and ckit (right panel) from virgin MECs. (E) qPCR showed Ly6a and Wnt4 enriched in the ML population and Elf5 in the LP population thus verifying the purity of each population. (F) mPINC was enriched in the luminal progenitors and alveolar progenitors. Data represent mean ±SD (n = 3). Target genes were normalized to Gapdh .

Article Snippet: Mammary epithelial cells were subsequently resuspended at a density of 1×10 7 cells/ml and stained with anti-mouse CD24 PE (Stem Cell Technologies, 1∶100), anti-CD49f FITC (Stem Cell Technologies, 1∶100), anti-mouse CD24 APC (Biolegend, 1∶100), anti-mouse CD29 Pacific Blue (BioLegend, 1∶100), anti-mouse CD14 FITC (eBiosiences, 1∶80) and anti-mouse ckit PE (Clone ACK4, Cedarlane Laboratories, 1∶50).

Techniques: Isolation, Expressing

Journal: Nutrients

Article Title: Identification and Purification of Novel Low-Molecular-Weight Lupine Allergens as Components for Personalized Diagnostics

doi: 10.3390/nu13020409

Figure Lengend Snippet: Immunoblot with ( A ) recombinant L. albus profilin and ( B ) with natural L. albus extract. ( A ) (1) Tris (buffer negative control) (2) negative serum; (3) peanut allergy, lupine-sensitized (P 11); (4) peanut allergy, lupine-sensitized (P 12); (5) suspected lupine allergy (P 14); (6) peanut allergy, lupine-sensitized (P 13); (7) peanut and lupine allergy (P 15); (8) peanut allergy, lupine-sensitized (P 16); (9) peanut allergy, lupine-sensitized (P 17). ( B ) The immunoblot was designed as an inhibition assay using natural L. albus extract and sera pre-incubated with different concentrations of recombinant L. albus profilin using one profilin-reactive serum (4). (1) Tris, (2) negative serum, (3) negative serum + 50 µg rProfilin, (4) serum 4 (P 12), (5) serum 4 + 5 µg rProfilin, (6) serum 4 + 50 µg rProfilin, (7) serum 4 + 500 µg lupine extract. Blocking was performed with Synblock; serum dilution was 1:20, dilution of the HRP-conjugated mouse-anti-human IgE Fc-antibody was 1:5000 (SouthernBiotech, Birmingham, AL, USA). Immunologically, the recombinant profilin performed similarly to the natural allergen. P-code refers to .

Article Snippet: Since the purification of the recombinant profilin via further chromatography steps (size exclusion chromatography and ion-exchange chromatography (source 15Q 4.6/100 PE, GE Healthcare, Uppsala, Sweden) were not successful, it was further purified via preparative SDS-PAGE.

Techniques: Western Blot, Recombinant, Negative Control, Inhibition, Incubation, Blocking Assay