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. 2019 Aug 13;11(11):967–978. doi: 10.1093/jmcb/mjz084

Figure 3.

Figure 3

Fusion of human trophoblast BeWo cells is negatively regulated by TTL. (A) Results of western blotting for BeWoGFP and BeWoTTL cells treated with DMSO or forskolin for 48 h using anti-tyr-α-tub, anti-detyr-α-tub, anti-Flag, or anti-GAPDH antibodies. To detect each type of modified α-tubulin, the samples were blotted separately. (B) The fusion index was determined contemporaneously with the samples harvested for western blotting. The total and assorted fusion indices are shown in the bar graph. **P < 0.01, *P < 0.05, n = 3. (C) For confocal microscopy, Alexa Fluor 488 and Alexa Fluor 555 were used to label E-cadherin and detyr-α-tub, respectively, in BeWoGFP or BeWoTTL cells treated with DMSO or forskolin for 48 h. Nuclei were stained with DAPI (blue). Scale bar, 20 μm. (D) TTL was stably knocked down using lentiviral shRNA in cells without any treatment or with forskolin treatment for 48 h. The fusion index was determined contemporaneously with the samples harvested for western blotting. **P < 0.01, *P < 0.05, n = 3.