Journal: Frontiers in Neurology

Article Title: Inhibition of Colony Stimulating Factor 1 Receptor Suppresses Neuroinflammation and Neonatal Hypoxic-Ischemic Brain Injury

doi: 10.3389/fneur.2021.607370

Figure Lengend Snippet: PLX3397 changes brain cytokine/chemokine profile in HI mice. (A) The total differentially expressed cytokines/chemokines in the brains of HI mice treated with PLX3397 or the vehicle. Brain tissues were collected at 48 h after HI induction and the protein levels were measured with a cytokine/chemokine profile ELISA array kit in which 111 factors were detected. n = 3 independent experiments. Each independent experiment needed 10 mice per group (5 males and 5 females). Unpaired two-tailed t -test. (B) The mRNA levels of representative differentially expressed factors related to regulation of chemotaxis (CCL2, CCL12), immune response (CD14, M-CSF, IL-7), and BBB integrity (ICAM-1, VEGF, and WISP-1). CX3CL1 was as a negative control that was not significantly changed in protein level between HI mice treated with PLX3397 or the vehicle (data not shown). Brain tissues were collected at 48 h after HI from HI mice treated with PLX3397 or the vehicle. n = 5 mice per group. Unpaired two-tailed t -test. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: Cytokines in the cerebral tissue lysates were measured using a mouse XL cytokine array kit (ARY028, R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test, Chemotaxis Assay, Negative Control

Journal:

Article Title: Intact Gram-Negative Helicobacter pylori , Helicobacter felis , and Helicobacter hepaticus Bacteria Activate Innate Immunity via Toll-Like Receptor 2 but Not Toll-Like Receptor 4

doi: 10.1128/IAI.72.11.6446-6454.2004

Figure Lengend Snippet: Murine macrophages are activated by whole H. pylori SS1 bacteria in a TLR2-dependent manner. (A) Wild-type mouse peritoneal exudate macrophages were stimulated with medium (left panel) or H. pylori SS1 bacteria (right panel). Following a 2-h incubation, RNA was extracted, labeled, and hybridized to a mouse SuperArray blot. The positions of inflammatory cytokine and cytokine receptor spots are indicated. (Bottom two rows) Negative controls included blanks and pUC18. Positive controls included the gene for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and housekeeping genes (those for cyclophilin A and actin). (B) Wild-type and TLR2 knockout (KO) mouse peritoneal exudate macrophages were stimulated with medium (None) or H. pylori SS1 bacteria (107 to 109 per well). Supernatants were harvested 18 h later, and IL-6 levels were determined by ELISA. (C) Wild-type, TLR2 knockout, or TLR4-deficient (def) ScN mouse peritoneal exudate macrophages were stimulated with medium alone or E. coli LPS (TLR4 ligand). Supernatants were harvested 18 h later, and IL-6 levels were determined by ELISA.

Article Snippet: Total mRNA (5 μg per blot) was reverse transcribed, labeled with biotin, and hybridized to a Mouse Inflammatory Cytokine SuperArray in accordance with the manufacturer's (SuperArray Inc., Frederick, Md.) instructions.

Techniques: Incubation, Labeling, Knock-Out, Enzyme-linked Immunosorbent Assay