Fig 1: MPC Assembly Is Affected upon Defects in TIM22 in Human Mitochondria(A) Topology of human MPC1 and MPC2.(B) Isolated mitochondria from control and patient fibroblast were analyzed by SDS-PAGE and western blotting.(C) HEK293T cells were treated with TIM22-specific small interfering RNA (siRNA) or non-targeting siRNA (NT). Isolated mitochondria were subjected to SDS-PAGE and western blotting.(D and E) [35S]-labeled (D) MPC1 and (E) MPC2 were imported into isolated mitochondria from HEK293T cells transfected either with non-targeting (NT) or TIM22-specific siRNA (TIM22KD).(F) Quantification of assembled MPC2 at 60 min (mean ± SEM, n = 3) (*p < 0.05, unpaired t test, two tails). aa, amino acid; ??, membrane potential.
Fig 2: MPC Subunits Are Imported along the Carrier Pathway(A) Isolated mitochondria from WT and tim22-14 cells grown for 15 h at the non-permissive temperature (37°C) were solubilized and analyzed by BN-PAGE and immunoblotting.(B and C) [35S]-labeled Mpc1 was imported into (B) WT and mpc1? mitochondria and (C) mpc1?, tim22-14/mpc1?, and tim10-2/mpc1? mitochondria.(D) [35S]-labeled Atp5 was imported into mpc1? and tim22-14/mpc1? mitochondria.(E) Membrane potential measurement of mpc1?, tim22-14/mpc1?, and tim10-2/mpc1? mitochondria.(F and G) [35S]-labeled Mpc3 was imported into (F) WT and tim22-14 mitochondria and (G) mpc1? and tim22-14/mpc1? mitochondria.(H) [35S]-labeled Mpc2 was imported into WT and tim22-14 mitochondria.(I) Purified mitochondria from WT and Tim22-downregulated (Tim22?) cells were analyzed by SDS-PAGE and western blotting.(J) Membrane potential measurement of WT and Tim22-downregulated mitochondria.(K and L) [35S]-labeled (K) Mpc2 and (L) Crc1 were imported into WT and Tim22-downregulated mitochondria. In all import experiments, analyses were carried out by BN-PAGE and digital autoradiography. ??, membrane potential.See also Figure S3.
Fig 3: Expression of MPC in HCC tissues and adjacent liver tissues. (A) Expression of MPC1 (left) and MPC2 (right) protein in HCC tissues vs. adjacent uninvolved tissue was analyzed by immunohistochemical staining. (B) Relative expression of MPC1 and MPC2 protein was calculated by the density mean. Error bars represent means ± standard deviation, ***P<0.001. (C) Western blot analysis of MPC1 and MPC2 in HCC samples and paired normal tissues. (D) Expression of MPC1 (left) and MPC2 (right) mRNA. Plotted bar represents the log2 mRNA expression ratio of non-cancerous tissue to tumor tissues. (E) Co-expression analysis for MPC1 in HCC vs. MPC2. Protein expression is represented by the density mean value (left) and mRNA expression is represented by ?Cq (right). MPC, mitochondrial pyruvate carrier; HCC, hepatocellular carcinoma.
Fig 4: Association between MPC protein expression and the OS and RFS rates of patients with hepatocellular carcinoma. (A) A significant difference in MPC1 expression was observed with regard to RFS and OS. (B) MPC2 expression did not differ significantly with regard to RFS and OS. MPC, mitochondrial pyruvate carrier; OS, overall survival; RFS, recurrence-free survival.
Supplier Page from Proteintech Group Inc for MPC2 antibody