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. 2021 Jul 14;12:685523. doi: 10.3389/fimmu.2021.685523

Figure 2.

Figure 2

Autophagy determination in LPS-induced HK-2 cells. (A) Representative western blot showing the Atg 5 and LC3II protein expression levels in HK-2 cells following LPS stimulation at various time-points. GAPDH was used as an internal reference. (B, C) densitometric analyses of Atg5 and LC3II protein expression. n = 4-5. (D) The autophagic flux in HK-2 cells was determined by cellular immunoassay following LPS stimulation at various time-points. The autophagy double-labeled adenovirus mRFP-GFP-LC3 was used to detect autophagic flux (magnification ×630 and scale bar = 10 µm). (E) The mean number of GFP and mRFP dots per cell. (F) The mean numbers of autophagosomes and autolysosomes per cell. * p < 0.05, ** p < 0.01 vs. the 0 h group, ## p < 0.01 vs. the 8 h group. The results are representative of at least three independent experiments. LPS, lipopolysaccharide; Atg5, autophagy-related protein 5; LC3II, Microtubule-associated protein 1A/1B-light chain 3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; mRFP, monomeric red fluorescence protein; GFP, Green fluorescent protein.