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. 2012 Jul 30;21(21):4645–4654. doi: 10.1093/hmg/dds306

Figure 2.

Figure 2.

Recombinant MBNL1 expression and localization in MBNL1-OE mice. (A) Western blots of tissues from FVB, 14685 and 14686 mice at 6 months of age. Recombinant MBNL1 and endogenous Mbnl1 are detected using A2764 (αMbnl1) primary and HRP-conjugated α-rabbit secondary antibodies. αFlag-HRP (M2) and α-mouse-HRP detect recombinant MBNL1 only. GAPDH was used as a loading control. (B) Western blots of whole embryo protein lysates show expression of recombinant and endogenous MBNL1/Mbnl1 for 14685 at embryonic day 11 (E11) and 14686 at E13. (C) Relative levels of recombinant (MBNL1) and endogenous (Mbnl1) protein in 14685 (n = 7) versus wild-type littermates (n = 3) and 14686 (n = 3) versus wild-type littermates (n = 3) were quantified and normalized to the alpha-tubulin loading control. Endogenous Mbnl1 levels in wild-type controls were set to 1 to calculate a fold-change in total MBNL1/Mbnl1 levels. *P < 0.05; **P < 0.005; ***P < 0.001. (D) Immunofluorescence of recombinant MBNL1 in 14685 gastrocnemius muscle using α-Flag (F7425 Sigma) and α-rabbit Alexa Fluor 488 (Jackson Immunoresearch) antibodies. No recombinant protein is detected in wild-type controls. DAPI stain was used to detect nuclei.