Journal: BMC Neuroscience

Article Title: Leucine-rich repeat kinase 2 (LRRK2) regulates α-synuclein clearance in microglia

doi: 10.1186/s12868-016-0315-2

Figure Lengend Snippet: Characterization of LRRK2-KO microglia. a Tomato lectin staining of LRRK2-KO and WT microglia. One hundred cells were counted per culture well. n = 3 culture wells per group. Scale bar 50 μm. b MTT assay for evaluating the viability of LRRK2-KO and WT microglia in the absence or presence of αSYN. The data represent percentage viability relative to culture day 0, and were assessed by ANOVA at each time point (3 h: F = 0.26, p = 0.852; 6 h: F = 1.26, p = 0.351; 10 h: F = 0.94, p = 0.465; 24 h: F = 3.61, p = 0.065; 48 h: F = 2.55, p = 0.129; and 96 h: F = 3.46, p = 0.071). n = 3 culture wells per group. c Phagocytosis assay using FITC-microbeads. Upper panel shows an image of microbead phagocytosis. Scale bar 50 μm. Lower panel shows the fluorescence intensity of microbeads taken up by microglia. Forty cells were analyzed per group. The data are expressed as mean ± SD and were assessed by Student’s t test. These experiments were carried out three times using independent primary microglia isolated from different mice, and a representative image and data are shown

Article Snippet: Primary microglia attached to cover slips were fixed with 4% paraformaldehyde for 30 min at room temperature and reacted with biotin-conjugated tomato lectin (Vector Laboratories) for 30 min at room temperature.

Techniques: Staining, MTT Assay, Phagocytosis Assay, Fluorescence, Isolation

Journal: PLoS Pathogens

Article Title: Modeling of the N-Glycosylated Transferrin Receptor Suggests How Transferrin Binding Can Occur within the Surface Coat of Trypanosoma brucei

doi: 10.1371/journal.ppat.1002618

Figure Lengend Snippet: Aliquots of purified T. brucei TfR (lanes 1, 2, 3, 6, 7, 10 and 11) and of bovine ribonuclease B, a positive control for ConA blotting (lanes 4 and 5), and bovine asialotransferrin, a positive control for ErCr lectin and ricin blotting (lanes 8, 9, 12 and 13), were separated by SDS-PAGE, transferred to nitrocellulose and subjected to blotting with anti-TfR antibody (lane 1), ConA (lanes 2–5), ErCr lectin (lanes 6–9) or ricin (lanes 10–13) in the absence (−) or presence (+) of the competing sugars α-methyl-mannose (lanes 3 and 5), lactose (lanes 7 and 9) or galactose and lactose (lanes 11 and 13). The positions of molecular weight markers are indicated for each group of blots.

Article Snippet: Ricin-biotin was used at a dilution of 1 in 3,000 (with or without 10 mg/ml galactose and 10 mg/ml lactose), tomato lectin-biotin conjugate diluted was used at a dilution of 1 in 10,000 (with or without chitin hydrolysate, Vector Laboratories, at a dilution of 1 in 10), ErCr lectin was used at a dilution of 1 in 3,000 (with or without 200 mM lactose).

Techniques: Purification, Positive Control, SDS Page, Molecular Weight