Figure 6. Swm regulation of expression and nuclear export of fucTA mRNA.

(A) Quantification of FucTA-Myc in control (gfp KD) and swm (swm KD) knockdown eyes. In this figure, all results shown in graphs are the mean ± SD, n = 2. (B) Expression of FucTA-Myc in the Golgi complex was reduced in swm^F14 mutants. FucTA-Myc (green) was well colocalized to the Golgi detected by anti-GM130 (red) in control eyes (arrows, +;fucTA-Myc^KI). In contrast, FucTA-Myc was hardly colocalized to the Golgi (arrowheads) in swm mutants (swm^F14; fucTA-Myc^KI). Scale bar: 10 µm. (C) In vivo binding of Swm and endogenous fucTA mRNA. Endogenous fucTA mRNA was immunoprecipitated with an anti-Flag antibody (M2) from BG2-c6 cells expressing Swm-Flag and detected by RT-PCR (bottom). Control experiments were performed without Swm-Flag expression (−) or by immunoprecipitation with a mouse IgG antibody (mIgG). (D) Decrease of fucTA mRNA in swm knockdown BG2-c6 cells. Levels of fucTA RNA in swm knockdown cells was normalized to that in control gfp knockdown cells, n = 3. ***p<0.005. (E) Decay of fucTA mRNA in swm knockdown BG2-c6 cells. Levels of fucTA mRNA at each time point after actinomycin D addition were normalized to those at time 0, n = 3. (F) Ratios of fucTA and actin (act5c) mRNA levels in nuclear vs. cytoplasmic fractions in swm knockdown BG2-c6 cells were normalized to those in control gfp knockdown cells. n = 4. (G) PolyA length of fucTA mRNA in swm knockdown BG2-c6 cells. PolyA of fucTA was amplified by PCR and detected by Southern blotting with a probe corresponding to fucTA.