FIGURE 3.

BCL2 interacting protein 3 (BNIP3) repression leads to activation of the NLRP3 (nucleotide‐binding oligomerization domain (NOD)‐, leucine‐rich repeat (LRR)‐, and pyrin domain‐containing protein 3) inflammasome and nuclear factor kappa B (NF‐κB)‐dependent inflammation in muscle. (a) Representative Western blot (WB) and quantification of NLRP3 inflammasome components and p65 protein in control (C) and BNIP3 knockdown (BNIP3 KD) myotubes (n = 6–9). (b) Interleukin 1β (IL‐1β) levels in culture medium from C and BNIP3 KD myotubes (n = 10). (c) Representative Western blot (WB) and quantification of BNIP3 protein expression in C and BNIP3 KD mouse gastrocnemius muscle (n = 5). (d) Representative WB and quantification of NLRP3 inflammasome components and p65 protein in C and BNIP3 KD gastrocnemius muscle (n = 10). (e) Representative immunofluorescence image and quantification of the co‐distribution between double‐stranded DNA (dsDNA), Toll‐like receptor 9 (TLR9), and lysosomal‐associated membrane protein 1 (LAMP1) in C and BNIP3 KD myotubes (n = 21 myotubes). (f) Representative WB and quantification of inflammasome components and p65 in C myotubes, and BNIP3 KD myotubes untreated or treated with TLR9 antagonist oligodeoxynucleotide (ODN) 2088 for 16 h (n = 6). Data are expressed as mean ± SE. *p < 0.05