Fig. 3

Pmpcb deficiency leads to impaired mitochondrial metabolism and function. A The fluorescence signal of the PmpcbDsRed protein partially co-located with MitoTracker® Green FM-labeled mitochondria in HeLa cells. B TEM analysis of mitochondrial structure in pmpcb−/− mutants at 72 hpf. Red arrows indicate the mitochondria and green staining indicate the endoplasmic reticulum (ER) in WT sibling and pmpcb−/− mutants, respectively. C-F WB analysis of mitochondrial-related functional proteins Clpp, Mtco1 (C, left), Hsp60 (D, left), and Lias (E) in pmpcb−/− mutants. Quantification of the relative levels of proteins Clpp, Mtco1 (C, right), Hsp60 (D, right), and Lias (F) in pmpcb−/− mutants and their WT siblings. G Images of 72 hpf head cells from the pmpcb−/− mutants and WT siblings staining by JC-1 output, and the images were gotten through fluorescence scanning confocal microscopy (G, left). G, right, Quantitative analysis of the ratio of red fluorescence intensity over green fluorescence. H ATP content in the pmpcb.−/− mutants and the WT siblings at 72 hpf. Each dot indicates one larva. I Transcriptional expression of the mature glial cell markers gfap and vimentin in the WTs, DMSO exposed WTs, and the CCCP exposed WTs at 24 hpf (I, left) and at 72 hpf (J, left), respectively. I, J, right, Calculation of gfap and vimentin expression levels in different samples. CCCP, a recognized mitochondrial uncoupler. I, J, lateral view, anterior to the left, and dorsal to the up. Each experiment was repeated at least three times, and a representative result is shown. Data are analyzed using GraphPad Prism 9.5 for t-test, and are presented as mean ± SD. ***P < 0.001. Scale bar, 200 μm (I, J), 5 μm (A, G), 1 μm (B)