Autophagosome Proteins LC3A, LC3B and LC3C Have Distinct Subcellular Distribution Kinetics and Expression in Cancer Cell Lines - Fig 2

<p><b>(A)</b> Confocal double immunofluorescence for LC3A (green) and LC3B (red) in various cell lines. Noted the LC3A accumulation in the perinuclear area, while LC3B is distributed throughout the cytoplasm (A1-7). LC3 aggregates, suggestive of small LC3A+ autophagosomes included in LC3B+ autophagosomes, are occasionally present (boxes in A1,2). Western blots confirm the nuclear presence of LC3A in the nuclei, mainly with the LC3A-II form (A8). <b>(B)</b> Nucleoli are stained with the MIB1/green (B1), the LC3B/red (B2), but not the LC3A/violet antibody (B3); LC3A specific ap1805a and of the LC3B specific 5F10 antibodies were used. This is confirmed in double immunostaining for LC3A/MIB1 (B4), LC3B/MIB1 (B5) and LC3A/LC3B (B6). Noted that LC3B stains the internal nucleolar areas, while MIB1 the peripheral. <b>(C)</b> Actinomycin D damages nucleoli and disrupts the nucleaolar LC3B and MIB1 localization (C1). Acidic conditions (pH = 6.5; C2) or hyperthermia (40°C; C3) do not abrogate the distribution of LC3B in the nucleoli nor of LC3A in the nuclei. Lamin is also used as a control to confirm the presence of nuclei only in the nuclear fraction.