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. 2011 Apr 1;7(4):401–411. doi: 10.4161/auto.7.4.14397

Figure 1.

Figure 1

Effects of ATRA on autophagy. (A) HL-60, NB4 and human PBMC cells were treated with ATRA (1 µM) for the indicated time with or without 3-methyladenine (“3-MA”, 10 mM), and then autophagy was assayed by quantification of the average number of LC3 puncta per cell. Representative images in HL-60 cells (48 h) are shown in the left part. Bar = 20 µm. (B) HL-60 cells were treated with ATRA (1 µM, 48 h) with or without 3-methyladenine (“3-MA”, 10 mM), pepstatin/E64D (“P/E”, 10 µg/ml) and LC3-I/II protein expression was assayed by western blot. Relative LC3-II expression is shown in the top part, “AU”: arbitrary unit (n = 3, *p = 0.024, ATRA + 3MA group versus ATRA group; p = 0.04, ATRA + 3MA group versus ATRA group). (C) HL-60 and NB4 cells were treated with ATRA (1 µM) or rapamycin (“Rapa”, 100 nM) for 24 h and phosphorylation of 4EBP1 (P-4EBP1) was assayed by western blot. (D) Ultrastructural features in HL-60 cells with or without ATRA (1 µM, 48 h) treatment. The number of autophagosomes observed under TEM was calculated (*p = 0.004, n = 3).