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. 2021 Mar 11;11:642271. doi: 10.3389/fcimb.2021.642271

Figure 4.

Figure 4

Over-expression of ATAT-HA induced the formation of an inclusion body-like structure. (A) Immunolocalization of ATAT-HA with rat monoclonal anti-HA antibodies in Dm28c pTcINDEX-GW ATAT-HA epimastigotes induced with 0.5 mg/ml tetracycline at different time points p.i. The yellow arrow indicates the inclusion body-like structure, the light blue arrow indicates the kinetoplast and the pink arrow indicates the nucleus. DAPI was used as nucleus and kinetoplast marker. Bar: 5 μm. (B) Transmission Electron Microscopy of Dm28c pTcINDEX-GW ATAT-HA epimastigotes induced with 0.5 μg/ml tetracycline for 48 h. ib, inclusion body-like structure. This structure was seen in close proximity with the endoplasmic reticulum (a, arrows) and was positioned close to the nucleus (n) and the kinetoplast (k) in the posterior part of the cell body (b) or more commonly at the anterior end, close to the kinetoplast and the basal body (c). The inclusions body is not surrounded by a membrane unit (d). bb, basal body; f, flagellum. The EDTA technique revealed ribosome rich regions in the nucleus, as the heterochromatin (ht) and granular domain of the nucleolus (white arrows), whereas the inclusion body-like structure (ib) lacks ribosomes and is less electrondense in relation to these regions. When comparing the inclusion body-like structure with organelles, as the nucleus (n) and the mitochondrion (m), it is clear that the first one is not surround by membrane units (e).