JCS040113 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Cldn10 variants stably transfected into MDCK-C7 cells. Confocal laser scanning micrographs of methanol-fixed cells (YFP-expressing cells, PFA-fixed) immunostained using the following antibodies: red, rabbit anti-Claudin-10 (Zymed, 1:100), Alexa Fluor 594 anti-rabbit (Zymed, 1:500); green, mouse anti-occludin (Zymed, 1:100), Alexa Fluor 488 anti-mouse (Zymed, 1:500), or YFP fluorescence, as applicable. Yellow staining in merged images indicates colocalization of claudin-10 and the tight junction marker occludin.

• Supplemental Figure S2 -

  Fig. S2. Transepithelial resistance and Na⁺ to Cl⁻ permeability ratio of transfected MDCK-C7 cell layers. (A) Transfection with Cldn10 caused transepithelial resistance (R^t) to decrease in all MDCK-C7 cell layers except those transfected with mouse Cldn10a (**P<0.01, mean ± s.e.m., n=4-41). Clones #1 are those shown in Fig. 5A, clones #2 are alternative, stably transfected clones for comparison. (B) Na⁺ over Cl⁻ permeability ratio (P_Na/P_Cl) was greatly increased in Cldn10b-transfected MDCK-C7 cell layers (**P<0.01, mean ±s.e.m., n=3-35). Clones #1 are those shown in Fig. 5B, clones #2 are alternative, stably transfected clones for comparison.

• Supplemental Figure S3 -

  Fig. S3. Western blot analysis of MDCK-C7 clones transfected with Cldn10 variants. Expression of endogenous claudins was determined in MDCK-C7 clones stably transfected with Cldn10 variants, which were used for electrophysiological studies. Expression was comparable for all claudins investigated (1-5, 7, 8) with the exception of claudin-1, which was downregulated in one clone transfected with human claudin-10b. As results in this clone did not qualitatively differ from results obtained from the clone transfected with mouse claudin-10b or from an alternative clone transfected with human claudin-10b, this downregulation was considered irrelevant for the present study.