Fig. 5

CG42724 influences TDP-43 production by regulating qualitatively and quantitatively the pattern and the nucleocytoplasmic export of TDP-43 mRNA transcripts. a Quantification of the TDP-43 mRNA steady-state levels by RT-QMPSF experiments in GMR-Gal4 > UAS-TDP-43_TDPBR and GMR-Gal4 > UAS-TDP-43_TDPBR, UY5237 flies. The graph represents mean ± SEM after normalization with Cyp1 (reference gene). Controls were arbitrarily set at 100 arbitrary units. The mRNA levels were compared between both genotypes by using Student’s t-test. ns: not significant. CG42724 expression did not significantly influence TDP-43 mRNA steady-state levels (n = 4, p = 0.055). b Total RNA from cytoplasmic and nuclear fractions were isolated from GMR > TDP-43_TDPBR or GMR > TDP-43_TDPBR, UY5237 transgenic flies. The graph represents mean ± SEM of TDP-43 mRNA levels detected by RT-QMPSF experiments, after normalization with Cyp1. Controls were arbitrarily set at 100 arbitrary units. TDP-43 expression levels were compared by using Student’s t–test. ns: not significant. CG42724 expression did not modulate global TDP-43 mRNA nucleocytoplasmic export (n = 3, p = 0.992). c Agarose gel electrophoresis of the PCR products in the 3’ RACE analysis. The 3’ RACE experiments were performed using RNA described in (a). Left: representative gel image (n > 10). Expression of CG42724 resulted in a qualitative distinct pattern. Right: schematic representation of the long and the short TDP-43_TDPBR mRNA variants detected in flies. d Quantification of the relative abundance of the TDP-43_TDPBR mRNA variants by 3′ RACE PCR amplification combined to QMPSF methodology, in GMR-Gal4 > UAS-TDP-43_TDPBR and GMR > TDP-43_TDPBR, UY5237 transgenic flies. The mRNA fractions from the cytoplasm and from whole cells (total RNA) were analyzed. Three fluorescent peaks corresponding to a short isoform, intermediate species and a long isoform were detected after separation by capillary electrophoresis (Additional file 7: Figure S5C). e The graph represents the ratio of the relative abundance of the short isoform/long isoform in each experimental condition presented in (d) (n = 4, GMR-Gal4 > UAS-TDP-43_TDPBR: p = 0.038, GMR > TDP-43_TDPBR, UY5237: p = 0.017)