Fig. 1.

Gal-9 depletion causes morphological and ciliogenesis defects. (A–C) A highly sensitive, electrochemiluminescence-based MSD technique was used to quantitate Gal-9 protein levels in retroviral shRNA-mediated knockdown cells. shRNA-susceptible canine and shRNA-resistant human Gal-9–EGFP constructs were stably expressed in MDCK cell lines to test the specificity of shRNA-mediated down-regulation. Samples were normalized to amounts of GAPDH. A one-tail, unpaired t test was used to generate P values. Error bars indicate SD. The data represent two mock-infected and shRNA cell pairs from three independent experimental groups. (D) Acetylated tubulin stains ciliary axoneme and basal body; ZO-1 reveals TJ areas. shRNA cells are enlarged, fusiform, and flat and show complete lack of cilium with occasional staining of the basal body (white arrowheads). (Scale bars: 10 μm.) The x–z view reveals TJs at varying heights.