JCS021485 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. The localization of claudin is similar in MDCK II and C7 cells. MDCK II (A,C,E,G,I,K) and C7 (B,D,F,H,J,L) cell lines were cultured for 4 days and processed for immunofluorescent analysis. The localization of claudins is similar in both cell lines, with the exception of claudin-2, which is expressed at such low levels in the C7 cells that it is difficult to visualize. Claudin-1 (C,D) and claudin-2 in MDCK II cells (E) colocalize with ZO-1; other claudins are more diffuse on the lateral membrane, including claudin-3 (G,H), claudin-4 (I,J) and claudin-7 (K,L). ZO-1 is included as a control. Bar: 20 µm (A,B).

• Supplemental Figure S2 -

  Fig. S2. Induction of claudin-2 and claudin-4 has little effect on the levels of endogenous claudins. Stable lines of MDCK II Tet-off cells were induced (i) or uninduced (u) to express claudin-2 (two separate clones, left panel) and claudin-4 (two separate clones, right panel) for 4 days, and total cell lysates were immunoblotted for claudin-1 (top panels), claudin-2 (second panels), claudin-3 (third panel), claudin-4 (fourth panel) and claudin-7 (bottom panel). Induction of claudin-2 was 4-5 fold and induction of claudin-4 was 3-7 fold compared with control values. Induction of claudin-2 resulted in a slight (20%) increase in the levels of claudin-1, whereas induction of claudin-4 resulted in a small (less than twofold) decrease in the levels of claudin-7. The levels of other claudins and occludin (data not shown) were unchanged after induction.