ELDR overexpression affects growth and cell cycle progression. (A-B) ELDR-overexpressing THP-1 lines ELDR-C1, ELDR-C3 and ELDR-overexpressing Mono-Mac-1 lines ELDR-C5 and ELDR-C7 reveal delayed growth measured by trypan blue exclusion when cultured in normal media (NM) compared with control cell lines established by transfection with EV controls. n = 3 biological replicates. (C-D) Growth curves of the ELDR-overexpressing THP-1 cell line (ELDR-C1) or EV control (EV1) and ELDR-overexpressing Mono-Mac-1 cell line (ELDR-C7) or EV control (EV2) in HS-5 conditioned media. n = 3 biological replicates. (E) Representative fluorescence-activated cell sorter (FACS) plots revealing EdU incorporation (y-axis) and DAPI staining (x-axis) of cells from the ELDR-overexpressing THP-1 cell line (ELDR-C1) or the EV control cell line (EV1) and ELDR-overexpressing Mono-Mac-1 cell line (ELDR-C7), or EV control (EV2) grown in HS-5 conditioned media. Cell cycle phase gates and percentage of cells in each gate are indicated on the plot. (F-G) Quantification of cells in G1, S, or G2/M phase of the indicated cell lines grown in NM or HS-5 conditioned media. n = 3 biological replicates. (H) Quantification of cells in G1, S, or G2/M phase from the ELDR-overexpressing THP-1 cell line (ELDR-C1^ctrl) or the EV control cell line (EV1^ctrl) vs ELDR deleted cell lines ELDR-C1^{del ELDR} or EV1^{del ELDR} grown in HS-5 conditioned media (del ELDR: CRISPR/Cas9 with gRNAs specific to ELDR; ctrl: CRISPR/Cas9 with nontargeting gRNAs). n = 3 biological replicates. Data are presented as mean ± SD. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. P values were calculated using 1-way ANOVA with Dunnett's multiple comparisons test (statistical analysis: EV1 vs ELDR-C1, ELDR-C3 for THP1, EV2 vs ELDR-C5, ELDR-C7 for Mono-Mac-1) (panels A-B); parametric unpaired t test (panels C-D); 1-way ANOVA with Holm-Šídák's for multiple comparisons test (panels F-H). ns, not significant.