Fig. 5: Pex5 KD-mediated PIS induces upd3 through JNK signaling.
a Expression of Mad, kay, Jra, yki in oenocytes of PromEGS>Pex5RNAi flies, n = 2 biological samples from 2 independent experiments. b P-JNK immunostaining of Pex5 KD oenocytes with (+RU, blue dots) or without (−RU, grey dots). Scale bar: 6.7 µm. c Quantification of b N = 6 biological replicates (8 nuclei per replicate). d Western blots showing the levels of P-JNK and JNK (arrows) in control (−RU) and Pex5 KD (+RU) oenocytes. e Quantification of d N = 2 independent biological samples. f Fluorescence imaging of JNK reporter TRE-DsRed from control (−RU) and oenocyte Pex5 KD (+RU). Scale bar = 20 μm. Quantification is shown on the right. N = 9 flies. g Expression of puc in oenocytes from flies with Pex5 kd oenocyte. N = 4 independent biological samples. h Expression of kay and puc in oenocytes dissected from flies with oenocyte-specific knockdown of ND-75 and Cat. N = 4 independent biological samples. i Expression of upd3 in oenocytes dissected from flies with Pex5 KD or Pex5; kay double KD. N = 4 biological samples. Ctrl genotype is UAS-Pex5RNAi/attP40; PromEGS-Gal4/+. + indicates wild type. j Representative M-mode of flies with oenocyte Pex5 KD, Pex5; upd3 double KD, or Pex5; kay double KD. k Arrhythmia of flies with oenocyte Pex5 KD, or Pex5; upd3 double KD, or Pex5; kay double KD, or Pex5; Jra double KD. Ctrl genotype is the same as i (nleft-right = 34, 30, 16, 17, 20, 16, 20, 13, 13, 14 flies). l Arrhythmia of flies with oenocyte upd3 KD (nleft-right = 20, 17). m Expression of IL-6 in human PEX1-G843D-PTS1 cells. N = 3 independent biological samples. n Western blots showing P-JNK and JNK in wild-type and human PEX1-G843D-PTS1 cells. o Quantification of Panel n. nJNK/tubulin = 2, nPJNK = 3 biological samples. Data are represented as mean ± SEM. P values are calculated using two-sided unpaired t-test (c, e–g, l, m), two-way ANOVA followed by Holm-sidak multiple comparisons (a, h, i, k, o) ns: not significant.
