primary antibodies against human mt 1 (mel1ar, n-20, sc13179)  (Santa Cruz Biotechnology)

Journal: Molecules

Article Title: JND4135, a New Type II TRK Inhibitor, Overcomes TRK xDFG and Other Mutation Resistance In Vitro and In Vivo

doi: 10.3390/molecules27196500

Figure Lengend Snippet: JND4135 displays TRK kinase inhibitory activities against both wild-type (WT) and resistant mutation. ( A ) structure of JND4135; and representative graphs of kinase inhibitory activities of JND4135 against TRKA ( B ), TRKB ( C ), TRKC ( D ) and TRKA-G667C ( E ). All kinase inhibitory assays included at least two independent experiments.

Article Snippet: Primary antibodies against phospho-TRKA/TRKB/TRKC (ab197071) and CDK4 (ab199728) were purchased from Abcam.

Techniques: Mutagenesis

Journal: Nucleic Acids Research

Article Title: Aberrant RNA processing contributes to the pathogenesis of mitochondrial diseases in trans -mitochondrial mouse model carrying mitochondrial tRNA Leu(UUR) with a pathogenic A2748G mutation

doi: 10.1093/nar/gkac699

Figure Lengend Snippet: Mitochondrial complex activities and morphology in mito-mice tRNA Leu(UUR)2748 . ( A ) Representative images of the levels of indicated subunits of Complexes I, II, IV, and V in livers of 10-month-old mice examined by western blotting. ND1, ND2, ND5 and CO1 are mtDNA-derived proteins, while others are nuclear DNA (nDNA)-derived. TOM20, VDAC and β-ACTIN were used as loading controls. ( B ) Native–PAGE gel images show the amount of each mitochondrial respiratory complexes (top) and the activity of Complexes I and IV (bottom) in kidney, liver, and brain tissues of 10-month-old mice in the gel. Quantification of Complex I activity ( C ) and Complex IV activity ( D ) in the kidney, liver, and brain (Each group, n = 3). Data are presented as the mean ± SD; * P < 0.05, ** P < 0.01 by Tukey–Kramer test. ( E ) Histochemical analysis of Complexes II and IV activities in heart and kidney from 10-month-old mice. Complex IV-positive (COX+) cells are stained in brown, and Complex IV-negative/Complex II-positive (COX-SDH+) cells are stained in blue. Note that kidney and heart of high % group mice contain COX-SDH + cells. Glomeruli (‘G’) of renal cells were not sufficiently stained owing to a scarcity of mitochondria. Scale bar = 100 μm. ( F ) Electron microscopy images of mitochondria in soleus muscle and liver from 10-month-old mice. The ‘L’ indicates lipid droplets. Scale bars = 1 μm. ( G ) The mitochondrial areas in soleus muscle and liver were analyzed by histogram and violin plot (each group has at least 200 mitochondria). ** P < 0.01, *** P < 0.001, **** P < 0.0001 by Kruskal–Wallis test .

Article Snippet: Proteins separated by using 8% or 12% SDS-PAGE gels were transferred to polyvinylidene difluoride (PVDF) membranes and blocked with PVDF blocking reagent for Can Get Signal (Toyobo) for 1 h. Membranes were then incubated with primary antibodies against MT-ND1 (1:1000; #ab181848; Abcam), MT-ND2 (1:1000; #19704-1-AP; Proteintech), MT-ND5 (1:1000; #PA5-36600; Thermo Fisher Scientific), NDUFB8 (1:1000; #ab110242; Abcam), NDUFS4 (1:1000; #ab139178; Abcam), SDHA (1:1000; #11998S; Cell Signaling Technology), MT-CO1 (1:1000; #ab14705; Abcam), COX4 (1:1000; #4850S; Cell Signaling Technology), ATP5A (1:1000; #ab14748; Abcam), VDAC (1:1000; #4866S; Cell Signaling Technology), TOM20 (1:1000; #sc-17764; Santa Cruz Biotechnology) or β-ACTIN (1:3000; #A1978; Sigma-Aldrich) overnight at 4°C [Can Get Signal immunoreaction enhancer solution 1 (Toyobo) was used for dilution].

Techniques: Western Blot, Derivative Assay, Clear Native PAGE, Activity Assay, Staining, Electron Microscopy