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. Author manuscript; available in PMC: 2018 Mar 1.
Published in final edited form as: Cell Microbiol. 2016 Oct 11;19(3):10.1111/cmi.12668. doi: 10.1111/cmi.12668

Figure 3. Infection with L. amazonensis disrupts paxillin-dependent adhesion complex formation.

Figure 3

(A) Effect of L. amazonensis infection on phosphorylated paxillin protein levels. BMDM were infected or not with L. amazonensis axenic amastigotes for 1 h. BMDM were washed to remove extracellular parasites, cultured at 34°C for another 24 h, fixed and stained with anti-phosphorylated paxillin antibodies. The arrows point to L. amazonensis-infected BMDM. On the right is shown the fluorescence intensity from 40 cells for each group quantified using Volocity software. ***, p < 0.001 (Student’s t test) compared with non-infected control. Bar: 30 μm. (B) Effect of L. amazonensis infection on total paxillin protein levels. BMDM were infected or not with axenic L. amazonensis amastigotes (La) for 1 h, washed and cultured at 34°C for another 24 h, followed by quantification of total paxillin by Western Blot. Anti-actin antibodies were used as loading controls. Phospho-paxillin/actin ratios were determined by densitometry. (C) Effect of L. amazonensis infection on phosphorylated paxillin protein levels. BMDM were infected or not with axenic L. amazonensis amastigotes (La) for 1 h, washed and cultured at 34°C for another 24 h, followed by quantification by Western Blot with anti-phospho-paxillin antibodies. Anti-actin antibodies were used as loading controls. Phospho-paxillin/actin ratios were determined by densitometry. (D) Effect of L. amazonensis infection on phosphorylated FAK protein levels. BMDM were infected or not with axenic L. amazonensis amastigotes (La) for 1 h, washed and cultured at 34°C for another 24 h, followed by quantification by Western Blot with anti-phospho-FAK antibodies. Anti-actin antibodies were used as loading controls. Phospho-FAK/actin ratios were determined by densitometry. The results shown in A–D are representative of three independent experiments. (E) BMDM were infected or not with axenic L. amazonensis amastigotes (La) for 1 h, washed and cultured at 34°C for another 24 h, followed by staining with phalloidin. The arrows point to L. amazonensis-infected BMDM. The images shown are representative of two independent experiments.