Skip to main content
Figure 6 | Veterinary Research

Figure 6

From: Mycoplasma bovis subverts autophagy to promote intracellular replication in bovine mammary epithelial cells cultured in vitro

Figure 6

M. bovis translocate into autophagosomes indicated by M. bovis co-localizing with GFP-LC3. A Representative images of M. bovis (Red) co-localizing with GFP-LC3 (Green) at 6 hpi. bMECs transfected by ad-GFP-LC3 was infected by M. bovis. Autophagy was enhanced by rapamycin (20 μg/mL) or starvation (Hanks balanced salt solution) after infection. Autophagy was inhibited by pre-incubating bMEC with 5 mM 3-methyladenine in full nutrition medium for 3 h prior to infection. The cells on coverslips were fixed by 4% paraformaldehyde, blocked by 3% with BSA in PBS and permeabilized with 0.2% Triton X-100 in PBS. The M. bovis in cells were immunostained with mouse anti-M. bovis antibody and Alexa Fluor-conjugated secondary antibody. NT: bMEC without treatment of rapamycin, Hanks balanced salt solution, 3-methyladenine or infection; Control: M. bovis-infected bMEC without treatment of rapamycin, HBSS, 3-methyladenine; 3-Ma: M. bovis-infected bMEC treated with 3-methyladenine; HBSS: M. bovis-infected bMEC treated with Hanks balanced salt solution; Rapamycin: M. bovis-infected bMEC treated with rapamycin. B Quantification of M. bovis co-localizing with GFP-LC3. Images were analyzed quantitatively with Image J software with the JaCoP plugin. Twenty cells for each sample and at least 60 cells in each group were used for statistical analyses. Data are mean  ±  SD of 3 independent experiments. Standard deviations of individual measurements are indicated as bars. *Compared to the control group (p  <  0.05).

Back to article page