Skip to main content
. Author manuscript; available in PMC: 2021 Feb 17.
Published in final edited form as: Cell Rep. 2021 Feb 2;34(5):108689. doi: 10.1016/j.celrep.2021.108689

Figure 2. Regulated mitochondrial depolarization and lysosomal acidification are critical for proper epidermal differentiation.

Figure 2.

(A) H&E staining of DMSO- and BafA1-treated epidermal cultures after 9 days, which highlights KH granules (white arrowhead) and retained nuclei in cornified layers (yellow arrowhead).

(B) WB of LC3 and early (Dsg1, K10) and late (Loricrin) markers of epidermal differentiation in lysates from DMSO- and BafA1-treated cultures.

(C) BafA1 impairs lysosomal acidification, EACC inhibits autophagosome-LYSO fusion, and CCCP depolarizes mitochondria.

(D) H&E staining of cultures treated with DMSO or EACC, highlighting KH granules (white arrowhead) and retained nuclei (yellow arrowhead).

(E) Quantification of retained nuclei per high-powered field (hpf) (mean ± SD, n = 17 fields, **p = 0.0002) and FLG IF (mean ± SD, n = 20 fields, ***p < 0.0001) in DMSO- and EACC-treated cultures.

(F) IF of FLG in DMSO- and EACC-treated cultures.

(G) H&E staining of cultures treated with DMSO or CCCP shows KH granules (white arrowhead) and retained nuclei (yellow arrowhead).

(H) Quantification of retained nuclei (mean ± SD, n = 57 fields, ***p < 0.0001) and FLG IF (mean ± SD, n = 8 fields, *p = 0.0286) in DMSO- versus CCCP-treated cultures.

(I) IF of FLG in DMSO- and CCCP-treated cultures. Dashed lines mark the bottom of the epidermis.

White scale bars, 10 μm; black scale bars, 50 μm.