Primary Cilia Blockage Promotes the Malignant Behaviors of Hepatocellular Carcinoma via Induction of Autophagy

Figure 3

Inhibition of primary ciliogenesis induces autophagic flux. (a) SMMC-7721 and Huh7 cells were transfected with si-IFT88 or si-NC (negative control) for 72 h. The expression of LC3 II/I and p62 was determined by western blot analysis; the value under the band is the ratio of the blot and normalized to control. (b) After transfection for 72 h, SMMC-7721 cells were assayed by TEM. Autolysosomes (the rounded vacuolar structures with only one membrane containing cytoplasmic contents) are marked by red arrows. Uncleared mitochondria marked by black arrows (×1500 or ×10000). (c–f) SMMC-7721 and Huh7 cells were stably infected with RFP-GFP-LC3 double fluorescence lentivirus and then transfected with si-IFT88 or si-NC for 72 h; representative fluorescence images of GFP-LC3 puncta (green, autophagosomes) and RFP-LC3 puncta (red, autolysosomes) (×400) as well as the overlay images were shown and fluorescence puncta were counted by inverted confocal microscope. .

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