Figure 5.
SERPINB5 rescues defects in epidermal differentiation in response to DPM1 loss. (a) Western blot showing SERPINB5-GFP expression in primary human keratinocytes that lack DPM1. GFP served as control. GAPDH used as internal loading control. (b) H&E staining of 3D-RHE from sgDPM1 expressing GFP control or SERPINB5-GFP 12 days after airlift. Immunostainings of CK10 and filaggrin were used as differentiation markers. Scale bar: 50 µm distance on H&E images and 10 µm on immunostaining images. Insets zoomed 2× of original image. White dashed line indicates insert membrane. Panel shows representatives from three biological replicates. (c–e) Quantification of corneal layer thickness, total epidermal thickness, and viable (non-corneal) epidermal layer thickness from three independent biological replicates. Student’s t test, unpaired. (f) Violin plot showing quantification of intercellular spaces within the epidermis in sgDPM1 GFP and sgDPM1 SERPINB5 GFP 3D-RHE. The area for defining intercellular spaces was set to be >50 µm2 to exclude shrinking artifacts. A minimum of 15 individual fields of view were used for analysis from three independent biological replicates. Unpaired Student’s t test. (g and h) Immunostaining and analysis of DSP intensity of sgDPM1 GFP and sgDPM1 SERPINB5-GFP 3D-RHEs. Panel shows representative of four biological replicates. Scale bar: 10 µm distance. Unpaired Student’s t test used to determine statistical significance (N = 4). (i and j) H&E staining of sgDSP 3D-RHEs 12 days after airlift. DSP staining shows depletion of DSP in sgDSP 3D-RHE. Immunostainings for CK10 and filaggrin used as differentiation markers. Scale bar: 50 µm distance on H&E images and 10 µm on immunostaining images. White dashed line indicates insert membrane. Panel shows representatives from three biological replicates. (k–m) Quantification of total epidermal thickness, viable (non-corneal) epidermal layer thickness, and corneal layer thickness from three independent biological replicates. Student’s t test, unpaired. (n) Violin plot showing quantification of intercellular spaces within the epidermis in sgNT1 and sgDSP 3D-RHE. A minimum of 15 individual fields of view were used for analysis from three independent biological replicates. Unpaired Student’s t test. Source data are available for this figure: SourceData F5.

SERPINB5 rescues defects in epidermal differentiation in response to DPM1 loss. (a) Western blot showing SERPINB5-GFP expression in primary human keratinocytes that lack DPM1. GFP served as control. GAPDH used as internal loading control. (b) H&E staining of 3D-RHE from sgDPM1 expressing GFP control or SERPINB5-GFP 12 days after airlift. Immunostainings of CK10 and filaggrin were used as differentiation markers. Scale bar: 50 µm distance on H&E images and 10 µm on immunostaining images. Insets zoomed 2× of original image. White dashed line indicates insert membrane. Panel shows representatives from three biological replicates. (c–e) Quantification of corneal layer thickness, total epidermal thickness, and viable (non-corneal) epidermal layer thickness from three independent biological replicates. Student’s t test, unpaired. (f) Violin plot showing quantification of intercellular spaces within the epidermis in sgDPM1 GFP and sgDPM1 SERPINB5 GFP 3D-RHE. The area for defining intercellular spaces was set to be >50 µm2 to exclude shrinking artifacts. A minimum of 15 individual fields of view were used for analysis from three independent biological replicates. Unpaired Student’s t test. (g and h) Immunostaining and analysis of DSP intensity of sgDPM1 GFP and sgDPM1 SERPINB5-GFP 3D-RHEs. Panel shows representative of four biological replicates. Scale bar: 10 µm distance. Unpaired Student’s t test used to determine statistical significance (N = 4). (i and j) H&E staining of sgDSP 3D-RHEs 12 days after airlift. DSP staining shows depletion of DSP in sgDSP 3D-RHE. Immunostainings for CK10 and filaggrin used as differentiation markers. Scale bar: 50 µm distance on H&E images and 10 µm on immunostaining images. White dashed line indicates insert membrane. Panel shows representatives from three biological replicates. (k–m) Quantification of total epidermal thickness, viable (non-corneal) epidermal layer thickness, and corneal layer thickness from three independent biological replicates. Student’s t test, unpaired. (n) Violin plot showing quantification of intercellular spaces within the epidermis in sgNT1 and sgDSP 3D-RHE. A minimum of 15 individual fields of view were used for analysis from three independent biological replicates. Unpaired Student’s t test. Source data are available for this figure: SourceData F5.

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