Fig. 2: Plekhg5-mediates the unconventional secretion of Sod1 via Rab26.
From: Plekhg5 controls the unconventional secretion of Sod1 by presynaptic secretory autophagy
A Western blots showing the knockdown of Plekhg5 in MNs (sh-RNA-1 = #1; sh-RNA-2 = #2). B Reduced Sod1 levels in the medium of Plekhg5-depleted MNs as shown by Western blot. C Quantifications of the Sod1 intensities upon knockdown of Plekhg5 in MNs. n = 7 biological replicates. One sample t-test, two-tailed. D Western blots showing reduced Sod1 levels in the medium of Plekhg5-depleted NSC34 cells. E Quantifications of the Sod1 intensities upon knockdown of Plekhg5 NSC34 cells. n = 3 biological replicates. One sample t-test, two-tailed. F Western blot showing the Sod1 levels in the somatodendritic and axonal compartments upon Plekhg5 knockdown. LE low exposure, HE high exposure. Quantifications of the Sod1 intensities in lysates (G) and media (H) of the somatodendritic and axonal compartments upon knockdown of Plekhg5 in MNs. n = 4 biological replicates. Two-Way ANOVA, Šídák’s multiple comparisons test. I Western blot showing the knockdown of Rab26 in MNs (sh-RNA-1 = #1; sh-RNA-2 = #2). J Western blots showing reduced Sod1 levels in the medium of Rab26-depleted MNs. K Quantifications of the Sod1 intensities upon knockdown of Rab26 in MNs. n = 4 biological replicates. One sample t-test, two-tailed. L Western blot of MN lysates showing the knockdown of Plekhg5 and simultaneous expression of Flag-Rab26-WT or Flag-Rab26-QL. M Expression of Flag-Rab26-QL restores the reduced Sod1 medium levels after Plekhg5 knockdown as shown by Western blot. N Quantifications of the Sod1 intensities upon knockdown of Plekhg5 and simultaneous expression of Flag-Rab26-WT or Flag-Rab26-QL in MNs. sh-Luc, n = 12; sh-Plekhg5-E, n = 12; sh-Plekhg5-E+Flag-Rab26-WT, n = 5, sh-Plekhg5-E+Flag-Rab26-QL, n = 12; biological replicates. One-way ANOVA; Tukey’s multiple comparisons test. Box bounds are defined by the 25th and 75th percentiles. Extending whiskers represent data points within 1.5 times the interquartile range from lower and upper quartiles. Lines and crosses denote the median and mean. Sod1 secretion was calculated as the ratio between the amount of Sod1 in the medium and in the lysate. Sod1 levels in the lysates were adjusted to Tuj1 (MNs) or Actin (NSC34 cells). The normalized Sod1 intensities were set to 1 in each experiment Data are mean ± SEM. Source data are provided as a Source Data file.
