Journal: Cancer Cell International

Article Title: Darunavir analog precursors target mitochondrial metabolism in multiple myeloma and CLL.

doi: 10.1186/s12935-025-04142-w

Figure Lengend Snippet: The HIV protease inhibitors and BnpM-NH 2 affect the respiratory chain function altering the the expression of complex II and IV proteins . ( A ) WB analysis of NDUFB8, SDHB, UQCRC2, MTCO2, ATP5A1, and Actin expression in MM1S, RPMI-8226, and HG3 cells treated with BupM-NH2 and BnpM-NH2 (30 µM) for 48 h was performed. The intensity of NDUFB8, SDHB, UQCRC2, MTCO2, ATP5A1, and Actin intensity normalized to actin were quantified and plotted as bar graphs. The results are presented as mean ± SD of at least three replicates. ( B ) ROS production and mitochondrial mass in MM and CLL cells treated with BupM-NH 2 and BnpM-NH 2 (30 µM) for 24 h were assessed using mitosox and mitotracker assays. FACS analysis was employed to detect median fluorescent intensity (MFI), which was reported on a bar graph. Data, presented as mean ± SD, were obtained from at least three independent experiments. Significant differences ( P ≤ 0.05) were determined by one-way ANOVA with Tukey’s post analysis

Article Snippet: MTCO2 (1:6000, 26 KDa) , Proteintech , Cat#: 55070-1-AP; RRID: AB_10859832.

Techniques: Expressing

Journal: Cancer Discovery

Article Title: Copper Drives Remodeling of Metabolic State and Progression of Clear Cell Renal Cell Carcinoma

doi: 10.1158/2159-8290.CD-24-0187

Figure Lengend Snippet: Cu-dependent activation of CuCOX promotes oxygen consumption, reorganization of respiratory supercomplexes, and nucleotide biosynthesis essential for Cu-dependent tumor growth. A–D, OCR from representative Seahorse mitochondrial stress tests in 786-O ( A and B ) and RCC4 ( C and D ) cells. Quantification of basal respiration, maximal respiration (post-FCCP injection), and respiration coupled to ATP production. P value from paired t test. E, Blue NativePAGE (BN-PAGE) analysis of respiratory complexes using digitonin permeabilized mitochondria isolated by anti-TOM22 immunopurification from Cu Hi vs. Cu Lo cells. Immunoblotting for indicated respiratory complex subunits. Red brackets indicate RSC. F, In-gel activity assay (IGA) for cytochrome c oxidase. RSCs are indicated with red brackets. G, Quantification of Cu Hi complex IV (COX) IGA relative to Cu Lo . P values are calculated by a one-sample t test. H, Western blot for COX subunits MT-CO1 and MT-CO2 and RSC assembly factor COX7A2L in mitochondrial lysates. I, BN-PAGE of mitochondria shows enrichment for COX7A2L in RSCs (red bracket) in Cu Hi cells. J, Quantification of COX7A2L in western blots of RSCs shown in M . P values calculated by one-sample t test. K, BN-PAGE of mitochondria shows enrichment for COX17, chaperone of Cu to CuCOX, in respiratory complexes from Cu Hi cells. L, Total CL content measured using mitochondrial lipidomics in 786-O and RCC4 cells. RA, relative abundance. M, Total CL content in xenografts formed by 786-O cells or in XP374d tumors in mice fed low and high Cu diet. N, Cu effects on labeling of 6-PDG, intermediate of oxidative branch of PPP, after 5 hours of incubation with [ 13 C 6 ]-glucose. O, Fractional enrichment of [ 13 C 5 ]- nucleotides labeled from [ 13 C 6 ]-glucose in a total pool of each nucleotide in Cu Lo and Cu Hi cells after 24 hours of incubation. P, Fractional enrichment of nucleotides labeled from [ 13 C 5 , 15 N 2 ]-glutamine in a total pool of each nucleotide in Cu Lo and Cu Hi cells after 24 hours of incubation. Q, Gross images of tumors formed by control 786-O cells expressing nontargeting (NT) or cells with COX4I1 knockdown in mice fed matched Cu Lo or Cu Hi diets. Scale bar, 1 cm. R, Weight of tumors shown in Q at collection. P values were calculated by one-way ANOVA with the Holm–Šidák posttest. S, Gross images of XP374d tumors in mice fed with a Cu Hi diet treated with IACS-10579 or vehicle (V). Scale bar, 1 cm. T, Volume of subcutaneous tumors at the indicated time points. U, The weight of tumors shown in S at collection. V, Representative images of staining for MT-CO2 in noninvasive and invasive orthotopic xenografts. K, kidney tissue; LD, lipid droplets; M, muscle; T, tumor tissue. The dashed line indicates the boundary between the tumor and kidney tissue. Scale bars, 200 μm. Means ± SEM are shown; P values were calculated from unpaired two-tailed t test unless indicated. See also Supplementary Fig. S2 and Supplementary Tables S3, S4A, and S4B.

Article Snippet: The following antibodies were used: Ki-67 (Roche Ventana, 05278384001, RRID: AB_2631262 or Santa Cruz Biotechnology, sc-23900, RRID: AB_627859) and MT-CO2 (1:100, Novus, NBP3-16283, RRID: AB_3599404).

Techniques: Activation Assay, Injection, Isolation, Immu-Puri, Western Blot, Activity Assay, Labeling, Incubation, Control, Expressing, Knockdown, Staining, Two Tailed Test