Fig. 1: OPA1 maintains mitochondrial and multiple organelle morphology in NP cells.

A Immunofluorescence staining for OPA1 and MitoTracker Red in primary NP cells transduced with lentivirally delivered control (ShCtrl) and Opa1 (ShOpa1 #1 and ShOpa1 #2) ShRNAs. The staining experiment was performed 2 times independently. Scale bar: Top rows- 15 μm, bottom row- 2 μm. A’ Western blot to confirm OPA1 knockdown in NP cells. A” Mitochondrial morphology and network analysis. B TEM images of ShCtrl and ShOpa1 transduced cells. Scale bar: Top row − 600 nm, bottom row − 200 nm C Mitochondrial number measurements from OPA1-deficient NP cells; 28-30 cells were measured from two independent experiments. D The mtDNA content in control and OPA1-deficient cells. Data are represented as box and whisker plots showing all data points with median and interquartile range and maximum and minimum values. Statistical significance was performed using One-way ANOVA or Kruskal-Wallis test with Sidak’s multiple comparisons test as appropriate (A’, C, D). E Staining for peroxisome marker PMP70, early endosome marker EEA1, and cis-Golgi marker GM130. Scale bar: 10 and 2 μm E’, E” Morphology analysis of PMP70, EEA1 shows altered morphology of peroxisomes and early endosomes in ShOpa1 cells. E”’ Quantification shows increased fragmented cis-Golgi in ShOpa1 transduced cells. 100-110 cells were measured for each of the organelle markers. Statistical significance was measured with a two-sided chi-square test. Source data are provided as a Source Data file.