Involvement of autophagy in hypoxia-BNIP3 signaling to promote epidermal keratinocyte migration

a Immunofluorescence staining of BNIP3, LC3, and Atg5 in normal unwounded skin (day 0) and in day 5, day 10, and day 15 wounded sections obtained from C57BL/6 mice showing upregulation of BNIP3 and activation of autophagy in migrating epidermis during normal wound re-epithelialization. Wounds were close to re-epithelialization on day 10 and fully re-epithelialized on day 15. Nuclei were stained with 4′, 6-diamidino-2-phenylindole (DAPI, blue). Scale bar？=？50？μm. Epi epidermis, Derm dermis. Narrow dotted line indicates the interface between epidermis and dermis or leading edge of migrating epidermis (day 5 and day 10). b–d Graphs indicate the relative fluorescence intensities as determined by ImageJ software. The results are represented by the mean？±？SEM (n？=？3). e The indicated wound specimens were lysed in lysis buffer and utilized for the detection of LC3, Atg5, Atg7, Atg16L1, Beclin-1, p-ULK1, and BNIP3. f The indicated wound specimens were lysed in lysis buffer and immunoprecipitated with anti-BNIP3 or anti-LC3 antibodies followed by immunoblotting with anti-LC3 or anti-BNIP3 antibody. *P？<？0.05 vs. day 0 grou