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Fig. 1 | Retrovirology

Fig. 1

From: Replication-competent HIV-1 in human alveolar macrophages and monocytes despite nucleotide pools with elevated dUTP

Fig. 1

Confirmation of in vivo AM phenotype and comparison of dTTP, dUTP nucleotide and UBER enzyme levels with that of MDM. A Fluorescence microscopy of AM stained with DAPI DNA stain (blue), CD68 cell surface marker (red) and pHrodo Green E. coli metabolic marker (green). An enlarged merged image is shown on the right. B AM cells in A were counted using ImageJ and the percent of DAPI stained cells that were also stained with the other two markers was calculated. C dUTP and dTTP levels in extracts from dividing HAP1 cells, non-dividing MDM, and AM cells as determined using a single nucleotide extension assay (SNE) (Additional file 1: Fig. S1). The in vitro differentiated MDM and the in vivo differentiated AM cells were from the same healthy donors. D dUTP/dTTP ratio in extracts from dividing HAP1 cells, non-dividing MDM, and AM cells. E Baseline mRNA expression levels of proteins involved in uracil base excision repair (UBER) and dNTP metabolism in AM and MDM cells. Expression levels were normalized to dividing HAP1 cells for comparison purposes. 18S ribosomal RNA was used as the calibration standard in all measurements. Error bars represents SD, the mean was calculated with three biological replicates for HAP1 cells, and from two healthy donors for the AM and MDM data

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