Some forms of childhood malnutrition and growth stunting may be preventable with vaccines WUSTLmedschool NatureComms
To assess the impact of LT on HNF4-mediated transcription, pTR104 carrying six copies of the HNF4 transcriptional response element upstream of a secreted) into confluent Caco-2 cells. Media were exchanged 24 h after transfection, and after an additional 24 h, baseline samples of media were removed from each well and stored at −80 °C. Wells were then treated with a heat-labile toxin overnight .
SMAD4-mediated transcription was assessed by transient transfection of Caco-2 cells with the SBE4-luc plasmid containing four copies of the SMAD4 transcriptional response element 5’-GTCTAGAC-3'Following treatment with overnight treatment with LT, cells were resuspended in 80 µl of EMEM media and mixed with an equal volume of ONEGLO EX Reagent for 20 min on an orbital shaker at 300 rpm, then read on the luminometer as above.
. Uptake of thiamin by its respective and distinct carrier-mediated mechanism was determined by subtracting uptake of [H]-thiamin in the presence of a high pharmacological concentration of unlabeled thiamin from uptake in their absence; all uptake data points were calculated relative to total protein content of the different preparations and presented as a percentage relative to simultaneously performed controls.
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