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. 2017 Apr 6;7:46161. doi: 10.1038/srep46161

Figure 3. Acidic pH environment induces autophagy in murine osteoblastic cell line MC3T3-E1.

Figure 3

(A) More autophagosomes were observed in the media of pH 6.4 with typical double-membrane structures surrounded with cytoplasm and mitochondria. (B)The expression of LC3-II and P62 in OBs as the treatment time in pH 6.4 is prolonged. (C) Densitometric analysis of LC3-II/LC3-I at 6 h, 12 h and 24 h. **P < 0.01 vs.24 h, *P < 0.05 vs.24 h. (D) Densitometric analysis of P62/β-actin at 6 h, 12 h and 24 h. *P < 0.05 vs.6 h. (E) The expression of LC3-II and P62 in OBs as the pH value decreased for 6 h. (F,G) Densitometric analysis of LC3-II/LC3-I and P62/β-actin with pH 6.4, 6.8 and 7.4 for 6 h. *P < 0.05 vs. pH 7.4. (H) The expression of LC3-II/LC3-I and P62 in OBs in pH 6.4 with CQ as the treatment time was prolonged. (I,J) Densitometric analysis of LC3-II/LC3-I and P62/β-actin at 6 h, 12 h and 24 h in pH 6.4. *P < 0.05 vs.24 h + CQ. (K) The expression of LC3-II/LC3-I and P62 in OBs as the pH value decreased for 6 h. (L,M) Densitometric analysis of LC3-II/LC3-I and P62/β-actin among 6.4 + CQ, 6.8 + CQ and 7.4 + CQ for 6 h. **P < 0.01 vs.pH 7.4 + CQ, *P < 0.05 vs. pH 7.4 + CQ. (N) Immunofluorescence staining of LC3 were labelled red and nuclei were stained with blue. Size bar = 100 μm. The brightness of red around nuclei increased with the decrease of pH value in media. (O) Quantified analysis of immunofluorescence staining of LC3. **P < 0.01 vs.pH 7.4, *P < 0.05 vs. pH 7.4. Values are the mean ± sd, *P < 0.05.**p < 0.01, in comparison to the control by one-way ANOVA.