Journal: Microbial pathogenesis

Article Title: Upregulation of haematopoetic cell kinase (Hck) activity by a secreted parasite effector protein (Ta9) drives proliferation of Theileria annulata-transformed leukocytes.

doi: 10.1016/j.micpath.2024.107252

Figure Lengend Snippet: Fig. 1. Confirmation of GFP expression in BoMacGFP and BoMacGFP−Ta9 cells. A, Bovine macrophages (BoMac) stably expressing GFP, or GFP-Ta9 were grown and fixed on coverslips. Expression of GFP in both cell lines was confirmed by immunofluorescent microscopy to reveal a cytoplasmic and membrane associated pattern for GFP-Ta9 that was distinct from the diffuse cytoplasmic and nuclear GFP pattern. The cells were also stained with a rat anti-Ta9 serum (shown in red) to further confirm high to lower level Ta9 expression only in BoMacGFP−Ta9, and not in BoMacGFP cells. Bar = 20 μm. B, Western blot of protein extracts prepared from BoMacGFP and BoMacGFP−Ta9 cells to estimate overall GFP expression in both groups. Expression of the 70 kDa Ta9-GFP protein was detectably lower compared to 25 kDa GFP in the transfected bovine macrophages. Anti-Tubulin antibody was used as a loading control.

Article Snippet: Secondary antibody for α-tubulin (Chicken anti-mouse IgG conjugated with HRP, Santa Cruz, sc-2954) was diluted in the same PBST-BSA solution (1:5000) and incubated with the membrane for 1 h at room temperature.

Techniques: Expressing, Stable Transfection, Microscopy, Membrane, Staining, Western Blot, Transfection, Control