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Figure 3 | BMC Cell Biology

Figure 3

From: Mig12, a novel Opitz syndrome gene product partner, is expressed in the embryonic ventral midline and co-operates with Mid1 to bundle and stabilize microtubules

Figure 3

Immunofluorescence analyses reveal co-localization of Mid1 and Mig12 within the cell. (A) Immunofluorescence analysis after transient expression of MGFP-Mig12 (upper panel), HA-Mig12 (middle panel) and untagged Mig12 (lower panel) in Cos7 cells, revealing a diffuse distribution of the protein, in both the nucleus and the cytoplasm. (B) Co-expression of both Mid1 and Mig12 leads to co-localization of the two proteins in cytoplasmic bundles. Standard fluorescence microscopy shows formation of bundles only in Mid1 (left panels) and Mig12 (right panel) co-expressing cells. The arrow indicates a single transfected cell where Mid1 shows the classical distribution along normal interphase microtubules. (C) The co-localization is confirmed by confocal microscopy analysis in which HA-Mid protein is visible as a red signal and MGFP-Mig12 protein as a green signal; co-localization is indicated as a yellow signal in merged images. (D) Co-localization is also observed using the HA-Mig12 construct (middle panels) together with either a Mid1 OS truncated mutant (GFP-Mid1 1331insA) or a Mid1 mutant (GFP-MidD) retaining the coiled-coil domain, both localized in cytoplasmic bodies. No co-localization is observed when HA-TRIM19/PML protein is co-expressed with GFP-Mig12. The right panels represent the merged images.

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