Figure 1. Identification of Tara as an... | Rockefeller University Press

$Figure 1. Identification of Tara as an AJ component, whose knockdown down-regulates the expression of E-cadherin. (A) Immunoblotting of the liver, bile canaliculi (BC), and the cell–cell junctional fraction (JF) with an anti-Tara antibody. Tara was enriched in the BC and JF. (B) Immunofluorescence micrographs of MDCK cells. Cells were costained for Tara with E-cadherin or ZO-1 to mark adherens or tight junctions, respectively. Top panels: Tara, green; E-cadherin, red. Bottom panels: Tara, green; ZO-1, red. Bar, 10 µm. Tara was colocalized with E-cadherin at cell–cell adherens junctions, at a position basolateral to the tight junctions. (C) Immunoblotting of Tara-KD and control MDCK cells (two clones) for cell–cell adhering junctional proteins. Control: mock-transfected MDCK cells. KD1 and KD2: Tara-KD MDCK cells (two clones). E-cad, E-cadherin; Cad-6, cadherin-6; β-Cat, β-catenin; p120, p120-catenin; Vin, vinculin; Occ, occludin; Dp, desmoplakin. Loading controls were prepared by probing portions of the same filter by anti–α-tubulin and the antibody for the respective experimental protein, except for Cad-6 and β-Cat, which were probed in the presence of the loading control anti–α-tub at the same time by mixture of antibodies. (D) Immunofluorescence micrographs of co-cultured Tara-KD and control mock-transfected MDCK cells stained for E-cadherin and cadherin-6. The E-cadherin signals were decreased in the Tara-KD cells. Note that the cadherin-6 signals were increased by Tara-KD. Tara, green; E-cadherin, red; cadherin-6, green (cy5-staining). Bar, 10 µm. (E) Immunofluorescence micrographs of co-cultured Tara-KD and control mock-transfected MDCK cells stained for p120- and β-catenins. Note that the immunofluorescence levels of the p120- and β-catenins at AJs were not changed by Tara-KD.$

Figure 1.

Identification of Tara as an AJ component, whose knockdown down-regulates the expression of E-cadherin. (A) Immunoblotting of the liver, bile canaliculi (BC), and the cell–cell junctional fraction (JF) with an anti-Tara antibody. Tara was enriched in the BC and JF. (B) Immunofluorescence micrographs of MDCK cells. Cells were costained for Tara with E-cadherin or ZO-1 to mark adherens or tight junctions, respectively. Top panels: Tara, green; E-cadherin, red. Bottom panels: Tara, green; ZO-1, red. Bar, 10 µm. Tara was colocalized with E-cadherin at cell–cell adherens junctions, at a position basolateral to the tight junctions. (C) Immunoblotting of Tara-KD and control MDCK cells (two clones) for cell–cell adhering junctional proteins. Control: mock-transfected MDCK cells. KD1 and KD2: Tara-KD MDCK cells (two clones). E-cad, E-cadherin; Cad-6, cadherin-6; β-Cat, β-catenin; p120, p120-catenin; Vin, vinculin; Occ, occludin; Dp, desmoplakin. Loading controls were prepared by probing portions of the same filter by anti–α-tubulin and the antibody for the respective experimental protein, except for Cad-6 and β-Cat, which were probed in the presence of the loading control anti–α-tub at the same time by mixture of antibodies. (D) Immunofluorescence micrographs of co-cultured Tara-KD and control mock-transfected MDCK cells stained for E-cadherin and cadherin-6. The E-cadherin signals were decreased in the Tara-KD cells. Note that the cadherin-6 signals were increased by Tara-KD. Tara, green; E-cadherin, red; cadherin-6, green (cy5-staining). Bar, 10 µm. (E) Immunofluorescence micrographs of co-cultured Tara-KD and control mock-transfected MDCK cells stained for p120- and β-catenins. Note that the immunofluorescence levels of the p120- and β-catenins at AJs were not changed by Tara-KD.

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