Computer models were established to predict tight junction protein dynamics (pink and cyan lines in A–L) and were compared with experimental data (red and blue symbols in A–L) for occludin (A–D), claudin-1 (E–H), and ZO-1 (I–L)

Small area FRAP (A, E, and I), large area FRAP (B, F, and J), tight junction FLIP (C, G, and K), and intracellular FLIP experiments (D, H, and L) are compared. The models at the left show the assumptions required for the simulation to fit the experimental data. Specific values are given in .<p><b>Copyright information:</b></p><p>Taken from "The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state"</p><p></p><p>The Journal of Cell Biology 2008;181(4):683-695.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2386107.</p><p></p

(A) EGFP-occludin, –claudin-1, –ZO-1, and –β-actin were studied by FRAP

High magnification images of tight junction segments before and at the indicated time points after photobleaching are shown in the left panels. Corresponding kymographs are shown at the right. (B) Quantitative analysis of FRAP from experiments similar to those shown in B ( = 7, 6, 8, and 6 for occludin, claudin-1, ZO-1, and β-actin, respectively). (C) The mobile fraction and of recovery for each protein were calculated from the recovery curves in B. Error bars represent SEM. Bars, 2 μm.<p><b>Copyright information:</b></p><p>Taken from "The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state"</p><p></p><p>The Journal of Cell Biology 2008;181(4):683-695.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2386107.</p><p></p

(A) EGFP-occludin–expressing cells within confluent monolayers were studied by FRAP after photobleaching elongated tight junction regions

Representative images before and at the indicated time points after photobleaching and the corresponding kymograph are shown. (B) The effect of continuous photobleaching of EGFP-occludin within a region of the tight junction is shown in representative images at the indicated times and in the corresponding kymograph. (A and B) Quantitative analysis of the individual sites indicated by the colored arrows is shown at the right. (C) Fluorescence of PA-GFP was activated in the white areas shown in the image collected during activation. Images collected at subsequent times show diffusion of activated PA-GFP–occludin to adjacent regions. The corresponding kymograph and quantitative analysis of the indicated activation region and adjacent region are shown. (D) The effect of continuous intracellular photobleaching of an EGFP-occludin–expressing cell within a confluent monolayer is shown. The kymographs and quantitative analyses show FLIP analysis of tight junction–associated EGFP-occludin within the indicated regions of photobleached and adjacent control cells. Bars: (A and B) 5 μm; (C and D) 10 μm.<p><b>Copyright information:</b></p><p>Taken from "The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state"</p><p></p><p>The Journal of Cell Biology 2008;181(4):683-695.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2386107.</p><p></p

(A) EGFP–claudin-1–expressing cells within confluent monolayers were studied by continuous photobleaching of a region of the tight junction

Representative images at indicated times and the corresponding kymograph show the effect on tight junction fluorescence. Quantitative analysis of the individual sites indicated by the colored arrows is shown at the right. (B) The effect of continuous intracellular photobleaching of an EGFP–claudin-1–expressing cell within a confluent monolayer is shown. The kymograph and quantitative analysis shows tight junction–associated EGFP–claudin-1 fluorescence within the indicated region of a photobleached cell before and at intervals after photobleaching. (C) High magnification images and corresponding kymograph of EGFP–claudin-1 are shown. The mobile fraction and of wild-type EGFP–claudin-1 and EGFP–claudin-1 are shown in the graph at the right. Bars: (A) 5 μm; (B) 10 μm; (C) 2 μm.<p><b>Copyright information:</b></p><p>Taken from "The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state"</p><p></p><p>The Journal of Cell Biology 2008;181(4):683-695.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2386107.</p><p></p

(A) EGFP–ZO-1–expressing cells within confluent monolayers were studied by FRAP after photobleaching elongated tight junction regions

Representative images before and at the indicated times after photobleaching and the corresponding kymograph are shown. (B) The effect of continuous photobleaching of EGFP–ZO-1 within a region of the tight junction is shown in representative images at the indicated times and in the corresponding kymograph. (A and B) Quantitative analysis of the individual sites indicated by the colored arrows is shown at the right. (C) The effect of continuous intracellular photobleaching of an EGFP–ZO-1–expressing cell within a confluent monolayer is shown. The kymograph and quantitative analysis show tight junction–associated EGFP–ZO-1 fluorescence within the indicated tight junction region of a photobleached cell. Bars: (A and B) 5 μm; (C) 10 μm.<p><b>Copyright information:</b></p><p>Taken from "The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state"</p><p></p><p>The Journal of Cell Biology 2008;181(4):683-695.</p><p>Published online 19 May 2008</p><p>PMCID:PMC2386107.</p><p></p