Figure S1. The expression and localization of Syndecan-4 (red) and Wnt5A (green) were examined in M93-047 cells using immunofluorescent confocal microscopy. Treatment of M93-047 cells with recombinant Klotho for 16 h shows that the decrease in Wnt5A expression in the presence of Klotho correlates with a decrease in the expression of Syndecan-4.

Figure S2. (A) The effect of Klotho on the localization and expression of Filamin A (green) was examined in M93-047 cells. Filamin A shows an irregular, diffuse pattern in the untreated cells, but becomes more regular and relocates to the periphery of the cells as Wnt5A expression (red) is reduced in the presence of Klotho. (B) G361 cells were treated with 200 nM control (CTRL) or Klotho (KL) siRNA for 48 or 72 h. Total RNA was isolated and real-time PCR analysis of Klotho expression was performed. Levels were normalized to 18S expression. (C) Knockdown of Klotho in G361 cells results in Filamin becoming expressed in a more diffuse pattern throughout the cells. (D) M2 cells were either left untransfected (UT) or transfected with either wildtype (WT) or Calpain resistant (CR) Filamin A (FLNA), and then subjected to a transwell invasion assay.

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