FIGURE 1:
CDHR2 KO cells exhibit aberrant cell morphologies and decreased apical junction markers. (A) Three DPC Control and CDHR2 KO CL4 cells stained for F-actin (magenta) and ZO-1 (green). (B) Cell area measured in Control cells (n = 384) and CDHR2 KO cells (n = 259) from three experimental replicates. (C) Cell elongation (max Feret/min Feret ratio) measured from the cells analyzed in B. (D) A total of 12 DPC Control and CDHR2 KO CACO-2BBE cells stained for F-actin (magenta) and ZO-1 (green). Zooms show area in the dashed boxes, in this case highlighting straight versus ruffled junctions in Control and CDHR2 KO, respectively. (E) Junctional straightness (see Materials and Methods) from Control and KO cell junctional segments where 1.0 is a straight line; n = 62 segments from three experimental replicates per condition. (F) 21 DPC Control and CDHR2 KO CACO-2BBE cells stained for F-actin (magenta) and NM2C (green). (G) Mean NM2C and (H) ZO-1 intensities measured in CACO-2BBE cells from two experimental replicates, 10 fields per replicate, 20 fields total. Break in Y axis represents level of background fluorescence. In G and H, points represent individual image fields with mean ± SD. Significance levels from unpaired t tests calculated for experimental replicates are also shown above each plot (*p < 0.05, **p < 0.005). Scale bars: 40 µm (A, D, F), 10 µm (D, Zooms).