Figure 7: P38 inhibition decreased autophagic signaling by sequestering P62 in AML12 cells. | Scientific Reports

Figure 7: P38 inhibition decreased autophagic signaling by sequestering P62 in AML12 cells.

From: Interferon regulatory factor-1 activates autophagy to aggravate hepatic ischemia-reperfusion injury via the P38/P62 pathway in mice

Figure 7

(a) Immunofluorescence and quantitative changes of P62 (green) in AML12 cells after exposure to AdGFP, AdIRF-1 or AdIRF-1 plus SB203580. The nucleus was counter-stained with DAPI (blue). Scale bar: 10 μm. (b) Quantitative changes of autophagic vacuoles imaged using TEM in AML12 cells treated as described in (a). Scale bar: 0.5 um. (c) Western blot analysis of P38, p-P38, LC3II and P62 levels in AML12 cells treated as described in (a). (d) Relative expression of P62 mRNA in AML12 cells treated as described in (a), determined using qRT-PCR and expressed as fold change over that of AdGFP. (e) AML12 cells were transfected with AdIRF-1 or P62 siRNA before being subjected to a simulated IR treatment. Confocal images of yellow (autophagosomes) and red (autolysosomes) puncta were collected from AML12 cells infected with GFP-RFP-LC3. (f) The data was quantified by counting the number of autophagosomes (yellow dots) and autolysosomes (red dots) per cell. (g) Western blot analysis of LC3II and P62 levels in AML12 cells treated as described in (e). **P < 0.01 and ***P < 0.001.

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