Journal: Cell Reports Medicine

Article Title: Urolithin A improves muscle strength, exercise performance, and biomarkers of mitochondrial health in a randomized trial in middle-aged adults

doi: 10.1016/j.xcrm.2022.100633

Figure Lengend Snippet: Urolithin A confers a proteomic signature of improved mitochondrial metabolism and mitophagy in human skeletal muscle (A) Venn diagram summarizing pathway enrichment analysis results from proteomics data in vastus lateralis skeletal muscle. Data represent upregulated pathways with an adjusted p value <0.1 in subjects treated with placebo or with UA at 500 and 1,000 mg for 4 months compared with baseline. (B and C) Dot plots showing top enriched pathways (WikiPathways 2019 Human), ranked by protein ratio, in the UA 500- (B) and UA 1,000 mg (C) groups from (A). Dot color and size indicate adjusted p value and protein count, respectively. Significant pathways for the placebo group were filtered out to identify treatment-specific pathways. (D) Western blot analysis of protein lysates from vastus lateralis skeletal-muscle biopsies in subjects treated as above. For each subject, both baseline (Pre) and end-of-the-treatment (Post) samples were run, and membranes were probed for phospho-Parkin, total Parkin, and the mitochondrial proteins ATP5A (belonging to the OXPHOS complex V), UQCRC2 (complex IV), SDHB (complex II), and NDUFB8 (complex I). Tubulin and VDAC were included as markers of total and mitochondrial protein abundance, respectively. Dashed line separates samples from individual subjects. (n = 6 Pre and Post, biologically independent samples). (E) Quantification of phospho-Parkin over Tubulin protein intensity from western blots (WBs) in (D) (n = 6). Two-sided, paired t-test. (F) Quantification of NDUFB8 (left) and SDHB (right) protein intensity, normalized over VDAC from WBs in (D) (n = 6). ∗p < 0.05; ∗∗p < 0.01; two-sided, paired t test.

Article Snippet: The following primary antibodies were incubated overnight diluted in blocking buffer: UBE2N (SantaCruz, sc-376470, 1:3000), Tubulin (Proteintech, 10004185, 1:3000), phospho-Parkin S65 (Cell Signaling, #36866, 1:1000), Parkin (Cell Signaling, #4211, 1:1000), OXPHOS Antibody Cocktail (Abcam, ab110413, 1:2000), VDAC (Proteintech, 10866-1-AP, 1:1000).

Techniques: Western Blot, Quantitative Proteomics

Journal: Cell Reports Medicine

Article Title: Urolithin A improves muscle strength, exercise performance, and biomarkers of mitochondrial health in a randomized trial in middle-aged adults

doi: 10.1016/j.xcrm.2022.100633

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were incubated overnight diluted in blocking buffer: UBE2N (SantaCruz, sc-376470, 1:3000), Tubulin (Proteintech, 10004185, 1:3000), phospho-Parkin S65 (Cell Signaling, #36866, 1:1000), Parkin (Cell Signaling, #4211, 1:1000), OXPHOS Antibody Cocktail (Abcam, ab110413, 1:2000), VDAC (Proteintech, 10866-1-AP, 1:1000).

Techniques: Recombinant, RNA Extraction, Software, Protease Inhibitor

Journal: Translational Psychiatry

Article Title: Mitochondrial deficits in human iPSC-derived neurons from patients with 22q11.2 deletion syndrome and schizophrenia

doi: 10.1038/s41398-019-0643-y

Figure Lengend Snippet: a , b Western blot of MRPL40, VDAC, SDHA, MT-ND1, cytochrome b, and COX1 from iPSC-derived neurons at differentiation day (DD) 21. In the MRPL40 +/mut , there is decreased expression of MRPL40, and the mitochondrial DNA-encoded proteins MT-ND1, cytochrome b, and COX1, but no differences in the nuclear-encoded proteins VDAC and SDHA. c Quantitative PCR of MRPL40 , VDAC , MT-ND1 , Cytochrome b and COX1 . In MRPL40 +/mut , there is a small but significant decrease in expression of MRPL40 , and no change in levels of the other transcripts. * = p < 0.05, NS—not significant. For a and b , MRPL40 ( N = 7), VDAC1 ( N = 5), SDHA ( N = 3), MT-ND1 ( N = 8), cytochrome b ( N = 6), and COX1 ( N = 5). For c , N = 3.

Article Snippet: Primer sequences: ACTIN (Assay ID: Hs01060665 _g1), MRPL40 (Assay ID: Hs00186843_m1), VDAC1 (Assay ID: Hs01019083 _m1), COX1 (Assay ID: Hs02596864_g1); cytochrome b: forward primer: 5′-AGTCCCACCCTCACACGATTCTTT-3′, reverse primer: 5′-AGTAAGCCGAGGGCGTCTTTGATT-3′; mt-ND1: forward primer: 5′-ATGGCCAACCTCCTACTCCTCATT-3′, reverse primer: 5′-TTATGGCGTCAGCGAAGGGTTG TA-3′.

Techniques: Western Blot, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction