Fig. 10

Expression and regulation of NLRP3 in human DMD myotubes. Primary cultures of human myocytes from C (control) and/or DMD subjects. a Differentiated myotubes were treated or not with 5 μg/ml ApN for 24 h. Gene expression of human NLRP3 and miR-711 was next quantified, normalized to TATA-box-binding protein (TBP), and the subsequent ratios were presented as relative expression compared to control conditions. Values are means ± SEM for five to six independent cultures (i.e. run at different times and, for each time, from a new vial of cryopreserved myoblasts) from five (NLRP3) or three (miR-711) different subjects in each C and DMD group. The subjects were always chosen at random. #P < 0.05, ###P < 0.001 vs C; *P < 0.05 for indicated conditions. b, c Differentiated myotubes from C or DMD subjects were challenged by an inflammatory stimulus (a combination of TNFα and interferon gamma (IFNγ)), then transfected or not with miR-711 mimic or anti-miR-711 (or their respective controls: Ctrl+ or Ctrl–), while being treated or not by ApN. mRNA levels of NLRP3 and TNFα were normalized to TBP and presented as the relative expression compared to the basal condition (i.e. no inflammation, no transfection or any other treatments) represented by the dotted line. Values are means ± SEM for five to nine independent cultures derived from five different subjects in each C and DMD group. *P < 0.05, **P < 0.01, ***P < 0.001 for indicated conditions