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Fig. 5 | BMC Cell Biology

Fig. 5

From: Lipopolysaccharide induces bacterial autophagy in epithelial keratinocytes of the gingival sulcus

Fig. 5

Autophagy induction in cultured keratinocytes by PgLPS. a Cell viability was determined using trypan blue exclusion. The graph shows the viability of HaCaT cells exposed different concentrations (0, 0.5, 1.0, 5.0, 10, or 50 μg/ml) of PgLPS for 24 h. Results are presented as a percentage of the trypan blue exclusion in untreated cells. Data represent mean values ± SD in quadruplicate. b Western blotting analysis of LC3 and beclin-1 (BECN1) expression in HaCaT cells treated with different concentration of PgLPS for 24 h. β-actin (ACTB) was used as a loading control. Qauntification values shown in part were the fold increases normalized to those of ACTB on day 0. Results are shown below the corresponding blots. All experiments were performed in quadruplicate. c-e Effect of 3-MA or PMB on induction of autophagy in PgLPS-stimulated HaCaT cells. HaCaT cells were treated with 10 μg/ml PgLPS with or without 10 mM 3-MA or 100 μg/ml PMB for24 h. c Immunocytochemical detection of LC3-positive autophagosomes (green) in keratinocytes. Bar = 50 μm. d Quantitative analysis of the percentage of LC3-positive cell ± SD from five independent studies. *Significantly different at P < 0.05 compared with HaCaT cells treated without PgLPS and treated with 3-MA or PMB. e Western blotting analysis of LC3 and BECN-1 expression. β-actin (ACTB) was used as a loading control. Band densities were presened as LC3 or BECN-1 expression fold-increases (normalized to ACTB) and compared with the results for untreated HaCaT cells. Quantification results are shown below the corresponding blots. All experiments were performed in quadruplicate

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