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. 2021 Sep 24;28(4):662–676. doi: 10.1158/1078-0432.CCR-21-2105

Figure 2.

Figure 2. Losartan inhibits in vitro monocyte migration elicited by canine and human OS cell secretion of CCL2. A, CCL2 secretion by human OS cells quantified via ELISA. B, CCL2 secretion by canine OS cells quantified via ELISA. C, THP-1 monocyte trans-well migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest, *, P = 0.01, ***, P < 0.001, ****, P < 0.0001. D, Primary human donor PBMC trans-well migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (**, P < 0.01; ****, P < 0.0001). E, Primary canine donor PBMC trans-well migration to canine OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (***, P < 0.001; ****, P < 0.0001). F, Correlation of log10 transformed mean CCL2 secretion (pg/mL) in both canine and human OS cell line conditioned media with log10 transformed mean human THP-1 monocyte, human PBMC, and canine PBMC trans-well migration. Spearman r = 0.64. G, Representative whole well images (10× magnification) and quantification of losartan inhibition of CCL2 directed canine peripheral blood monocyte migration (n = 4 canine donors). Crystal violet staining. Data represent mean ± SEM of each donor, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05). H, Losartan inhibition of THP-1 monocyte migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). I, Losartan inhibition of human PBMC migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05; **, P < 0.01; ***, P < 0.001). J, Losartan inhibition of canine PBMC migration to canine OS cell line conditioned media, n = 5 independent canine donors. All THP-1, human PBMC, and canine PBMC trans-well migration assays were performed in technical triplicate or quadruplicate and are representative of a minimum of two different canine or human blood donors or three independent experiments (THP-1 cells).

Losartan inhibits in vitro monocyte migration elicited by canine and human OS cell secretion of CCL2. A, CCL2 secretion by human OS cells quantified via ELISA. B, CCL2 secretion by canine OS cells quantified via ELISA. C, THP-1 monocyte trans-well migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest, *, P = 0.01, ***, P < 0.001, ****, P < 0.0001. D, Primary human donor PBMC trans-well migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (**, P < 0.01; ****, P < 0.0001). E, Primary canine donor PBMC trans-well migration to canine OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (***, P < 0.001; ****, P < 0.0001). F, Correlation of log10 transformed mean CCL2 secretion (pg/mL) in both canine and human OS cell line conditioned media with log10 transformed mean human THP-1 monocyte, human PBMC, and canine PBMC trans-well migration. Spearman r = 0.64. G, Representative whole well images (10× magnification) and quantification of losartan inhibition of CCL2 directed canine peripheral blood monocyte migration (n = 4 canine donors). Crystal violet staining. Data represent mean ± SEM of each donor, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05). H, Losartan inhibition of THP-1 monocyte migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). I, Losartan inhibition of human PBMC migration to human OS cell line conditioned media. Data represent mean ± SD, analyzed by one-way ANOVA with Tukey posttest (*, P < 0.05; **, P < 0.01; ***, P < 0.001). J, Losartan inhibition of canine PBMC migration to canine OS cell line conditioned media, n = 5 independent canine donors. All THP-1, human PBMC, and canine PBMC trans-well migration assays were performed in technical triplicate or quadruplicate and are representative of a minimum of two different canine or human blood donors or three independent experiments (THP-1 cells).