Fig 1: Host protein expression and cellular distribution.(A) HNRNPR cellular distribution appears to be different in uninfected/early stage infected cells (U/E; blue outlines) than in late stage infected cells (L; white outlines). (B) MBOAT7 expression appears greater in late stage infected cells than in uninfected/early stage infected cells. (C–D) Plots of cellular Gag-immunofluorescence (IF) and unspliced (US) HIV RNA-fluorescence in situ hybridization (FISH) intensities for cells analyzed for HNRNPR (C) and MBOAT7 (D). Cells prior to the inflection points in each plot are termed ‘early cells’ as they are either uninfected or at stages of HIV replication prior to late gene expression (i.e., high amounts of Gag in the cytoplasm). Cells after the inflection point are termed ‘late cells’ as they express high, IF-detectable levels of Gag in the cytoplasm. (E–F) For each cell in the early and late cell sub-groups, the intensity of US HIV RNA vs the intensity of HNRNPR (E) and MBOAT7 (F) is plotted. Linear or polynomial regressions (R2) are fit to each early and late sub-group and the Pearson’s correlation coefficient (R) calculated for linear regressions. This demonstrates the extent of correlation between US HIV RNA expression and the expression of each host protein. (G–J) A Student’s t-test is applied to determine if the host protein intensities in early cells are significantly different from those in late cells in the nucleus and the cytoplasm. (K) The percent change in the median expression for all host proteins with early vs late p-values<0.05. Calculations were made for total cell, nuclear, and cytoplasmic differences. (L–M) Total cellular US HIV RNA intensities vs host protein nuclear to cytoplasmic (nuc/cyto) or nuclear plus perinuclear (nuc+peri)/cyto ratios. This demonstrates the extent of correlation of host protein cellular distribution with US HIV RNA expression. (N–O) A Student’s t-test measures significant differences in the cellular distribution between the early and late cells for HNRNPR and MBOAT7. (P) The percent change in the median nuc/cyto or nuc+peri/cyto ratio for all host proteins with p-values<0.05. Purple indicates the late cells have a higher proportion of the host protein in the cytoplasm than do the early cells. Green indicates the late cells have a higher proportion in the nucleus.Figure 5—source data 1.Quantitation of expression and distribution changes of host proteins in early and late HIV infection.