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. 2019 Apr 10;30(5):824–839. doi: 10.1681/ASN.2018090941

Figure 7.

Figure 7.

Thrombospondin type 1 domain–containing 7A (THSD7A) expression augments podocyte adhesion and slows motility. (A) Adhesion assay of THSD7A-expressing human podocytes in comparison with mock-transduced control cells to collagen type IV–coated 96-well plates. Adhering cells were quantified by a custom-made ELISA to β-actin. Values were calculated as percentage absorbance to the 30-minute timepoint, and they are depicted as the mean±SEM of pooled data from seven independent experiments with eight biologic replicates. *P>0.05 (between the time courses; two-way ANOVA, Bonferroni multiple comparison); **P>0.01 (between the time courses; two-way ANOVA, Bonferroni multiple comparison); §§P<0.01 (between the time courses; two-way ANOVA, Bonferroni multiple comparison). (B) Detachment assay of THSD7A-expressing human podocytes in comparison with mock-transduced control cells differentiated for 10 days on collagen type IV–coated 96-well plates. Podocyte detachment was obtained by incubation with 10 mM EDTA and quantified by a custom-made ELISA to β-actin. Values were calculated as percentage absorbance to 2.5 minutes of EDTA incubation (at which time no cell detachment has occurred yet), and they are depicted as the mean±SEM of pooled data from five independent experiments with eight biologic replicates. ***P<0.001 (between the time courses; two-way ANOVA, Bonferroni multiple comparison); §P<0.01 (between the time courses; two-way ANOVA, Bonferroni multiple comparison). (C) Migration assay of THSD7A-expressing and mock control human podocytes differentiated on collagen type IV–coated six-well plates for 10 days after the placement of a scratch at 0 hours (demarcated by the blue dotted line). After 21 hours, cells were fixed, and phase contrast micrographs were taken. The area of cells that had migrated into the wound (demarcated by the red line) was analyzed by ImageJ. The graph exhibits the filled area of scratch in percentage of mock. hTHSD7A, human thrombospondin type 1 domain–containing 7A. ****P<0.001 (two-tailed t test).