Table 1.
Primers used for real-time PCR experiments to detect the expression of various genes in the TA, SOL and DIA muscles of PBS- and LPS-injected mice.
Figure 1.
Representative electron microscopy images (A,B,C) of normal sub-sarcolemmal and intermyofibrillar mitochondria in SOL, DIA and TA muscles, respectively, of control mice.
Mit indicates mitochondria. T indicates triad. Representative electron microscopy images (D,E,F,) of mitochondrial morphological abnormalities in SOL, DIA and TA muscles of LPS-treated mice. Swollen mitochondrion (D, black arrow), mitochondrion densely packed with protein aggregates (E, black arrow), mitochondria with extensive loss of internal cristae (F, white arrows). Representative electron microscopy images of mitochondria with myelin-like structures (G, white arrow), extensive vacuole formation (H) and dilated sarcoplasmic reticula (I) in TA muscles of LPS-treated mice.
Figure 2.
Means ± SEM of proportion of mitochondria with ultrastructure morphology scores between 0 to 4 as percent of total counted mitochondria in TA, SOL and DIA of control and LPS-treated mice.
Mitochondria were assigned a score ranging from 0 to 4 based on integrity of matrix, internal cristae and membrane and EM density. N = 5 per group. *P<0.05, as compared to control group.
Figure 3.
Representative example (A) and means ± SEM of mitochondrial Ca2+ retention capacity of the TA, SOL and DIA muscles of control and LPS-treated mice (B).
*P<0.05, as compared to control group. N = 6 per group.
Figure 4.
Means ± SE of mRNA expressions of transcription factor regulators of nuclear-encoded mitochondrial proteins (Nrf1, Nrf2, Pgc1α, Pgc1β), regulators of mitochondrial DNA transcription (Tfam, Tfb1 and Tfb2) and cytochrome c oxidase subunits Cox4i2 and Cox1 in the TA, SOL and DIA muscles of LPS-treated mice.
N = 6 per group. Values expressed as fold change relative to control group. *P<0.05, as compared to control group.
Figure 5.
Means ± SEM of mRNA expressions of BclXL (anti-apoptotic) and Bim, Bad, Bax and Bnip3 (pro-apoptotic) genes in the TA, SOL and DIA muscles of LPS-treated mice (A).
Representative immunoblots of Bnip3 protein in the TA, SOL and DIA muscles of control and LPS-treated mice (B). Means ± SEM of Bnip3 protein (60 kDa)(C). N = 6 per group. Values expressed as fold change relative to control group. *P<0.05, as compared to control group.
Figure 6.
Representative electron microscopy images of autophagosome formation in skeletal muscles of LPS-treated mice.
Double-membrane autophagosomes in close proximity containing one (A) or more than one mitochondrion (B) in TA muscle. Double-membrane autophagosomes containing damaged mitochondria in the SOL (C, white arrow) and DIA (D, black arrows). Four-membrane autophagosome (C, black arrow) is also shown. Autophagosomes containing intermyofibrillar mitochondria and lipid droplets in TA (E and F, respectively) were also evident.
Figure 7.
Means ± SEM of mRNA expressions of autophagy-related genes in the TA, SOL and DIA muscles of LPS-treated mice.
N = 6 per group. Values expressed as fold change relative to control group. *P<0.05, as compared to control group.
Figure 8.
Representative immunoblots of Lc3b, Beclin1, p62 and tubulin (A) and Pi3kC3 and Atg5-Atg12 complex in the TA, SOL and DIA muscles of control and LPS-treated mice.
Means ± SEM of autophagy-related protein optical densities (B) and the ratio of Lc3b-II/Lc3b-I protein optical densities (C) in the TA, SOL and DIA muscles of LPS-treated mice. N = 6 per group. Values expressed as percent relative to control group. *P<0.05, as compared to control group.
Figure 9.
Representative immunoblots of IκBα, phospho-p65 (RelA) and tubulin proteins (A) and means ± SEM of optical densities of IκBα and phospho-p65 (n = 6, B) in the TA muscles of WT mice after 3, 6 and 24 h of LPS injection.
Representative immunoblots of IκBα and tubulin proteins in the TA muscles of WT and MISR mice (C). Means ± SEM of mRNA levels of autophagy-related genes in the TA muscle of WT and MISR mice measured after 24 h of LPS injection and normalized as fold change from control (n = 6, D). Representative immunoblots (E) and means ± SEM of Lc3b-II optical density (n = 6, F) of TA muscle measured after 24 h of LPS injection. Data are shown as fold changes from control values. *P<0.05, as compared to control group. #P<0.05 compared with wild type mice.