You were working with mouse mitochondrial proteins, and as a routine, you have isolated muscle tissues and homogenized it in ice-cold conditions. Following homogenization, purification was carried out by differential velocity centrifugation, equilibrium density gradient centrifugation, and finally, you have collected the fractions. But you have failed to detect any mitochondrial marker enzyme citrate synthase activity in the fractions. When you visualized the samples under the microscopes, you observed broken organelle debris due to severe proteolytic degradation. Which of the following statement may justify the cause of this result?

URGENT Cisplatin si a compound, hypothesized to target mammalian Cytochrome C, localized ni the mitochondrial electron transport chain. You have a culture of hepatocellular HepG2 cells, that you have treated with Cisplatin at 2 uM for 3 hours.Based on what you have learned from fluorescent microscopy in this tutorial, design an experimentto investigate whether this compound affects the mitochondrial ETC, in HepG2 cells. You have access to all the fluorophores, fluorescent probes and fluorescent proteins shown in the table below, alongwith technologies to label antibodies and create fluorescent protein expressing cell lines. In your experimental planning include:1) 2) 3)4) 5)Experimental justification and principle.How you will determine mitochondrial function with your selected fluorophores.Complementary fluorophores to perform mitochondrial colocalization. Draw an image of your excitation and emission spectra.The positive and negative controls in your experiment. The expected results…

The intermediates A, B, C, D, E, and F all occur inthe same biochemical pathway. G is the product of thepathway, and mutants 1 through 7 are all G−, meaningthat they cannot produce substance G. The followingtable shows which intermediates will promote growthin each of the mutants. Arrange the intermediates inorder of their occurrence in the pathway, and indicatethe step in the pathway at which each mutant strain isblocked. A + in the table indicates that the strain willgrow if given that substance, an O means lack of growth.SupplementsMutant A B C D E F G1 + + + + + O +2 O O O O O O +3 O + + O + O +4 O + O O + O +5 + + + O + O +6 + + + + + + +7 O O O O + O +

The enzyme dihydrofolate reductase (DHFR) normally resides in the cytosol, andDHFR can be imported into mitochondria by appending a mitochondrial signalsequence. However, when this modified DHFR is incubated with methotrexate, whichis a substrate analog that binds tightly to the active site, the modified DHFR is nolonger imported.a) Propose an explanation for why methotrexate prevents import of DHFR intomitochondria.b) Suppose that DHFR were modified instead by appending a nuclear localizationsignal. Would you expect methotrexate to prevent transport of this modified DHFRinto the nucleus? Why or why not?