Fig. 1

Cystinosis podocytes display impaired mitochondrial metabolism and a fragmented mitochondrial network. a. Intracellular metabolite levels in CTNS?/?#1 and CTNS?/?#2 normalized to WT podocytes measured by LC?MS (n?=?3 biological experiments, n???2 technical replicates). b. Cystine concentrations in WT, CTNS?/?#1, and CTNS?/?#2, expressed as nmol mg?1 protein. (n???6 biological experiments, n?=?2 technical replicates). Statistical analysis: Dunnett?s one-way ANOVA. (c, d) c. Citrate, ?-ketoglutarate, fumarate, and malate levels in CTNS?/?#1 and CTNS?/?#2, normalized to WT podocytes and protein (mg?1), d. ATP, NAD, and NADH levels in CTNS?/?#1 and CTNS?/?#2, normalized to WT podocytes and protein (mg?1). (n?=?3 biological experiments, n???2 technical replicates). Statistical analysis: linear mixed model for each metabolite. AU?=?arbitrary units. LOD?=?below the limit of detection. Each dot represents a technical replicate. (e, f) Mitochondrial fragmentation in cells expressing the mitochondrial roGFP2 redox sensor. Left panel: quantification of mitochondrial fragmentation (number of fragments normalized to cell area) in e. CTNS?/?#1 and CTNS?/?#2 normalized to WT podocytes; f. shCTNS#1 and shCTNS#2 normalized to shCTR. (n?=?4 biological experiments, n???4 technical replicates). Statistical analysis: linear mixed model. Each dot represents an individual cell measurement. Right panels: representative images of mitochondrial networks. Scale bar:?20 ?m. Unless noted, each dot represents a biological experiment