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. 2017 Jan 9;8(2):205–221. doi: 10.1080/19491034.2017.1279775

Figure 5.

Figure 5.

The relationship between rRNA synthesis and the autophagy inducer LC3-II in the nucleoli of ETO-treated cells and a possible role of p16INK4A in channelling of selected nucleoli: (A) NT control – RPA194 ‘punctuates’ the sites of transcription on the background of weak LC3-II presence in the nucleoli, while scarce LC3II foci in cytoplasm indicate the basic autophagy; (B) ETO - the number of cytoplasmic autophagosomes is much increased, while the nucleolar pattern is heterogeneous. In some cells RPA194 remains at the sites of transcription (FC and enlarged on insertion), while in other, it has left them and coalesced nearby (arrowheads) indicating cessation of rDNA transcription, while the presence of LC3-II in such nucleoli is increased; (C) – in some cells with coalesced RPA194 it appears to be substituted in the nucleolar sites with bright LC3-II foci; (D) p16 is seen in the selected nucleolus with ‘torulus” (red- arrowed, while a normal nucleolus is white-arrowed) and perinuclearly (enframed); (E) P16 is in the nuclear channel and perinuclearly (enframed); (F, G) p16 is found in 3 hollow DNA and FIB free nucleoli with a ‘torulus’, while the fourth arrowed nucleolus contains both FIB and DNA; (H-J) NoAs, nuclear fragmentation, and cell death: 2 ETO cells, one is engulfing the other (RGB image). The engulfed dead cell contains 2 nuclear NoAs with extremely high concentration of LC3-II, the cytoplasm is very poor with LC3-II, while the DAPI-stained nucleus is disintegrated (RGB image); (I) The enlarged fragment of the 2 NoAs in the engulfed cell, (I) in the red channel for LC3-II, (J) green channel for FIB; (K) An example of the FIB-NoA (arrow) releasing from a cell nucleus together with the DNA material in the breast cancer cell after neoadjuvant therapy (doxorubicin and paclitaxel) of the patient (from the operational material). The patchy staining for lamin B1 suggests its impairment. The tissue specimens were collected after the patients’ informed consent was obtained in accordance with the regulations of Committee of Medical Ethics of Latvia. Bars = 10 µm.