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problem
An experiment utilising live *Escherichia coli* (EC) and killed *E. coli* (KEC) bacteria was set up. KECs were generated by treating EC with streptomycin or gentamycin, and both ECs and KECs were stored at 4°C prior to use. Firstly, ECs were subjected to stable isotope labelling by amino acids in culture (SILAC) or grown in a culture medium only containing 13C-labelled glucose (13CLG) before being killed to generate SILAC-KECs or 13CLG-KECs, respectively. Both types of KECs were then added to a culture of bone marrow derived macrophages (BMDMs) grown in (A) transwell chambers or (B) 2D cell culture flasks before being analysed via liquid chromatography combined with mass spectrometry (LC-MS). It was found that the SILAC and 13CLG signals were only present in BMDMs grown in 2D cell culture flasks, but not transwell chambers. Additionally, SILAC signal in cell culture flasks was abrogated upon treatment with concanamycin A. These results were also found to be applicable to several other types of both gram positive and gram negative bacteria. Then, BMDMs were stimulated with 13CLGEC or 13CLGKECs until the internalised bacteria had been largely degraded, and metabolomics analysis was performed using LC-MS. It was found that KEC-treated macrophages displayed higher levels of glutathione, glutathione disulfide and increased levels of itaconate production than the EC-treated macrophages. BMDMs were then cultured in cell culture media deprived of amino acids before being incubated with either 13CLGEC or 13CLGKECs. It was found that EC-treated macrophages displayed lower cell viability than the KEC-treated macrophages, which was only rescued upon addition of a cocktail containing both essential and non-essential amino acids. What is the higher cell viability in KEC-treated macrophages under conditions of amino acid deprivation attributed to? Think step by step and solve the problem below. At the end of your response, write your final answer on a new line starting with “FINAL ANSWER”. It should be an answer to the question such as providing a number, mathematical expression, formula, or entity name, without any extra commentary or providing multiple answer attempts.
subject
biology
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