Journal: Cellular and molecular gastroenterology and hepatology

Article Title: ANKRD22 Drives Rapid Proliferation of Lgr5 + Cells and Acts as a Promising Therapeutic Target in Gastric Mucosal Injury.

doi: 10.1016/j.jcmgh.2021.06.020

Figure Lengend Snippet: Figure 5. ANKRD22 deletion reduces the inflammatory response by inhibiting activation of the macrophages after gastric mucosal injury. (A and B) Ankrd22 knockout decreased the number of CD45þ cells in damaged mouse gastric mucosa detected by FCM. Rat IgG2b was used as an isotype control (n ¼ 4). (C and D) Effect of Ankrd22 knockout on the expression profiles of inflammatory factors in damaged mouse gastric mucosa detected by Luminex assay (n ¼ 4). (E) Ankrd22 knockout reduced IL1a and TNF-a in mouse damaged gastric mucosa detected by ELISA (n ¼ 3). (A–E) Ankrd22þ/þ and Ankrd22-/- mice were examined 24 hours after intragastric administration of EtOH/HCl or saline solution (Ctrl). (F) Expression levels of ANKRD22 in the macro- phages activated by different concentrations of IFN-g. THP-1 and RAW264.7 macrophages were stimulated with 0, 50, 80, or 100 ng/mL IFN-g for 24 hours. Subsequently, the expression of ANKRD22 was determined by qRT-PCR. The data from the 0 ng/mL group were normalized to 1.0. TUBB and Tubb were used as internal references. (G) Expression levels of Ankrd22 in the activated mouse macrophages detected by qRT-PCR. Data of the PBS-treated group (Ctrl) were normalized to 1.0. Tubb was used as an internal reference. (H and I) Determination of IL1a and TNF-a in supernatant of activated macrophages from Ankrd22þ/þ or Ankrd22-/- mice by ELISA. (J and K) Effects of Ankrd22 knockout on the percentages of CD86þ and CD206þ cells in activated Ankrd22þ/þ and Ankrd22-/- mouse macrophages detected by FCM. Rat IgG2b was used as an isotype control (n ¼ 3). (L and M) Determination of IL12 (p70) and IL10 in the supernatant of activated macrophages from Ankrd22þ/þ or Ankrd22-/- mice by ELISA. (E–M) Macrophages were stimulated with 50 ng/mL IFN-g or 100 ng/mL LPS or PBS (Ctrl) for 24 hours. The data were analyzed by the Student t test and are presented as means ± SD. *P < .05, *P < .01, and ***P < .001. APC-A, Allophycocyanin Area; FITC-A, Fluorescein Isothiocyanate Area; FSC-A, Forward Scatter Area.

Article Snippet: According to the manufacturer’s instructions, the cytokine levels of IL1a, TNF-a, IL12, and IL10 in the samples were determined using ELISA kits (Proteintech).

Techniques: Activation Assay, Knock-Out, Control, Expressing, Luminex, Enzyme-linked Immunosorbent Assay, Saline, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Novel Cell Preservation Technique to Extend Bovine In Vitro White Blood Cell Viability

doi: 10.1371/journal.pone.0140046

Figure Lengend Snippet: Change in sample to positive ratios of interferon gamma production over time (days). On each day, this change was determined by comparing the use of a stimulant (pokeweed mitogen, PWM; concanavalin A, ConA; interleukin-12 p40 potentiated ConA, ConA+IL-12 p40; or IL-12 p40 alone) versus no stimulation (phosphate buffered saline) for white blood cells unpreserved or preserved with a transport medium (SCSR-T, NonInvasive Technologies) supplemented in whole blood samples from ten healthy cows.

Article Snippet: Stimulation agents for IFN-γ production and release analysis in this study included the mitogen PWM (Sigma-Aldrich, St. Louis, MO, USA), the mitogen ConA (Sigma-Aldrich, St. Louis, MO, USA), recombinant bovine IL-12 p40 (Kingfisher Biotech, St. Paul, MN, USA), and a combination of ConA and IL-12 p40.

Techniques: Saline

Journal: PLoS ONE

Article Title: Novel Cell Preservation Technique to Extend Bovine In Vitro White Blood Cell Viability

doi: 10.1371/journal.pone.0140046

Figure Lengend Snippet: Results from day 8 of the sampling period are shown for whole blood samples, either preserved (panel a.) with a transport medium (TM; SCSR-T, NonInvasive Technologies) or unpreserved (panel b.), that were collected from ten healthy cows. Arrows identify areas representing viable and nonviable cells, with the percentage of cells measured labelled per quadrant. Representative samples shown were samples stimulated with interleukin-12 p40 potentiated concanavalinA (ConA+IL-12).

Article Snippet: Stimulation agents for IFN-γ production and release analysis in this study included the mitogen PWM (Sigma-Aldrich, St. Louis, MO, USA), the mitogen ConA (Sigma-Aldrich, St. Louis, MO, USA), recombinant bovine IL-12 p40 (Kingfisher Biotech, St. Paul, MN, USA), and a combination of ConA and IL-12 p40.

Techniques: Sampling