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. 2019 Jul 9;9(17):5065–5084. doi: 10.7150/thno.34014

Figure 8.

Figure 8

Overexpression of SPRR family members cause abnormal cell junctions. (A) The lentivirus vectors and mSprr1b[ORF045957] were applied to mouse skin. After 3 days, H&E staining was performed. The expression and distribution of K6 were determined by immunohistochemistry. (B) The expression and distribution of ZO-1, E-cadherin and desmoplakin were determined semiquantitatively by immunofluorescence (C); n = 8; *p<0.05. (D) Primary keratinocytes were isolated from P1 WT mice, cultured in low-calcium conditions, and then infected with vector or msprr1b[ORF]. After 3 days, the proliferation of primary keratinocytes was detected using a Cell-Light EdU Apollo 567 In Vitro Kit. (E) The number of EdU-positive cells per 200X field of view from eight independent experiments, n = 8; *p<0.05. Primary keratinocytes were infected with vector or msprr1b[ORF], grown to confluency in low-calcium medium and then incubated in high-calcium medium for 12 h. Cells were stained for ZO-1 (F), E-cadherin (G) and desmoplakin (H). (I) Vector-infected keratinocytes and msprr1b[ORF]-infected keratinocytes were grown in low-calcium medium, and WB of cell lysates was performed with anti-ZO-1, E-cadherin and desmoplakin antibodies. Actin was used as a loading control. The experiments were repeated at least eight times; *p<0.05.