Fig. 3: Decreased GATD3A contributes to mitochondrial dysfunction and promotes FLS senescence. | Nature Communications

Fig. 3: Decreased GATD3A contributes to mitochondrial dysfunction and promotes FLS senescence.

From: GATD3A-deficiency-induced mitochondrial dysfunction facilitates senescence of fibroblast-like synoviocytes and osteoarthritis progression

Fig. 3

a Volcano plot of RNA-Seq data. b Western blotting analysis of GATD3A in synovium and FLSs (whole cell lysate) from normal and OA patients. c Quantitative PCR analysis of GATD3A in FLSs (n  =  25 Nor, n  =  33 OA). d Pearson’s correlation between GATD3A expression and CDKN2A expression in OA-FLSs (n  =  33). e Representative images of immunofluorescence of GATD3A in synovium. n = 15 independent biological samples. Scale bar: 100 µm. f Western blotting localization of GATD3A. n  =  3 independent biological replicates. g Fluorescent analysis of GATD3A localization, with signal intensity profiles along a line and Pearson’s correlation analysis (n  =  5 TOMM20, n  =  6 ATP5A and GRP75). Scale bar: 5 µm. Mitotracker staining (n = 10 ShNC, n = 12 ShGATD3A, n = 13 Vec, n = 11 GATD3A) (h), TEM (n = 16 ShNC and ShGATD3A, n = 8 Vec, n = 9 GATD3A) (i) were conducted. Scale bar: 40 µm for Mitotracker staining, 1 µm for TEM. OCR (n = 4 ShNC and ShGATD3A, n  =  5 Vec and GATD3A) (j) and PER (n = 5 ShNC and ShGATD3A, n = 6 Vec, n = 4 GATD3A) (k) assays of Nor-FLSs or OA-FLSs after different treatments. l, m Quantification of ATP content in FLSs after different treatments. n  =  6 independent biological replicates. Western blotting analysis (n  =  3 independent biological replicates) (n), SA-β-Gal staining (n = 6 ShNC and ShGATD3A, n  =  8 Vec and GATD3A) (o) and immunofluorescence (n  =  10 ShNC and ShGATD3A, n  =  13 Vec and GATD3A) (p) of FLSs after different treatments. Scale bar: 100 µm for SA-β-Gal staining, 50 µm for immunofluorescence. q, r Elisa assay to detect the pro-inflammatory cytokines. n  =  6 independent biological replicates. s Western blotting analysis of chondrocytes co-cultured with FLSs after different treatments. n  =  3 independent biological replicates. All data were presented as the means ± SD. P values were determined by two-tailed unpaired Student’s t-test or one-way ANOVA with Tukey’s multiple-comparisons (g). P < 0.05 was considered statistically significant. Source data are provided as a Source Data file.

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