Analyses of pERM and total ERM levels after knockdown of Clic5, Clic5a or Clic5b. (A) Immunoblot analyses and respective quantification reveals significant decreased pERM levels in whole protein lysates from 1dpf old embryos that were injected with SB-MO clic5 (2 ng) or TB-MO clic5b (2 ng) compared to TB-MO clic5a (3 ng) or Co-MO (3 ng) injected embryos. (B) Immunoblot analyses and respective quantification of total ERM levels reveals no significant difference in whole protein lysates from 1dpf old embryos that were injected with SB-MO clic5 (2 ng), TB-MO clic5a (3 ng) or TB-MO clic5b (2 ng) compared with Co-MO (3 ng) injected embryos. (C,D) Double whole-mount immunostaining and respective quantification of 1dpf old zebrafish embryos using anti-pERM (green) and anti-acetylated Tubulin (magenta) antibodies reveals significant reduced pERM levels in the pronephric tubules of embryos injected with SB-MO clic5 (2 ng) or TB-MO clic5b (2 ng) compared to TB-MO clic5a (3 ng) and Co-MO (3 ng) injected embryos; Arbitrary Unit (A.U.). Scale bar: 10 µm.
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