Fig. 9. Knockdown of mLgl induces formation of abnormal cell aggregates without obvious lumens during MDCK cell cystogenesis. (A) mLgl kd cells (#1) or control cells were suspended in collagen gel and cultured for the indicated time. Then cysts were fixed and stained with rhodamine-phalloidin and anti-E-cadherin antibody. Cell nuclei were stained with TOPRO3. Notice that mLgl knockdown affected the actin distribution during cystogenesis, leading to the formation of cell aggregates without lumens. (B) Quantification of abnormal cell aggregates in total cysts at day 7 is shown. Values are the mean (±s.d.) of three independent experiments. Notice that about 80% of cysts of both mLgl kd cells (#1 and #2) formed abnormal cell aggregates. (C) mLgl kd (#1) or control cells were suspended in collagen gel and cultured for 7 days, and then were fixed and stained with anti-PAR-6ß and anti-E-cadherin antibodies. Notice that the distribution of PAR-6ß to the inner surfaces of the cysts was affected in cysts of mLgl kd cells. Each photograph represents the confocal single x-y section. Bars, 20 µm.