Fig. 4: Impaired peroxisomal import in oenocytes induces upd3 and promotes cardiac arrhythmia. | Nature Communications

Fig. 4: Impaired peroxisomal import in oenocytes induces upd3 and promotes cardiac arrhythmia.

From: Impaired peroxisomal import in Drosophila oenocytes causes cardiac dysfunction by inducing upd3 as a peroxikine

Fig. 4

a Schematic diagram showing the key genes involved in peroxisomal import and membrane assembly. b Arrhythmia index of oenocyte-specific knockdown of mitochondrial complex I subunit ND-75 (two independent RNAi lines), mitochondrial Mn superoxide dismutase Sod2, peroxisomal import factors (Pex5, Pex1, Pex14), and peroxisomal membrane assembly factor (Pex19), (nleft-right = 14, 15, 14, 11, 16, 17, 18, 18 flies). Oenocyte-specific GeneSwitch driver (PromEGS-Gal4) was used. c Arrhythmia index of oenocyte-specific knockdown of peroxisomal matrix enzymes, Dhap-at, ADPS, Acox57D-d, Acox57D-p, Prx5 (nleft-right = 17, 18, 18, 18, 18, 17, 21, 20, 18, 16, 17, 14, 17 flies). d QRT-PCR analysis showing relative mRNA levels of upd3 from dissected oenocytes of Pex5, Pex1, Cat, Pex19 and ND-75 KD flies. Mito: Mitochondrion. N = 4 biological samples from 2 independent experiments. e QRT-PCR analysis showing relative mRNA levels of upd1, upd2, upd3 from dissected oenocytes of oenocyte-specific Pex5 KD flies. N = 3 biological samples, from 2 independent experiments. f Representative images of Stat92E immunostaining of heart tissues dissected from flies with (+RU) or without (−RU) oenocyte-specific Pex5 KD. Arrows indicate cardiomyocyte nuclei. White boxes indicate the regions shown in the right insets. Scale bar: 20 µm (inset: 5 µm). g Quantification on the number of Stat92E punctae around cardiomyocyte nuclei of oenocyte-specific Pex5 KD flies (blue dots). Dot plot shows the quantifications of 6 biological replicates, 3-4 selected regions of interest (ROIs) per replicate. h Representative images of Pmp70 immunostaining in fly tissues. Arrows indicate cardiomyocyte and pericardial cell nuclei. Scale bar: 6.7 µm. i Quantification of Pmp70-positive peroxisomes per region of interest. Dot plot shows the quantifications of 6 biological replicates, 4 ROIs per replicate. j Arrhythmia of flies with Pex5 KD in either oenocytes (PromEGS-Gal4) or fat body/gut (S106GS-Gal4) (nleft-right = 19, 22, 13, 17 flies). Data are represented as mean ± SEM. P values are calculated using two-sided unpaired t-test (g), one-way ANOVA followed (i), or two-way ANOVA (b–e, j) by Holm-Sidak multiple comparisons. ns: not significant.

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