Figure 5.

Effect of 1,25(OH)₂D₃ (50 nM) on HBFCs apoptosis using flow cytometry (A–G). HBFCs have been measured for Annexin V-FITC or 7AAD content and assigned to different cell populations by drawing gates, as demonstrated in each panel: Q1 (necrotic cells), Q2 (late apoptotic cells), Q3 (early apoptotic cells) and Q4 (living intact cells). TGF-β1-treated NHBFCs (A), 1,25(OH)₂D₃-TGF-β1-treated NHBFCs (B), TGF-β1-treated NHBFCs vs 1,25(OH)₂D₃-TGF-β1-treated NHBFCs, overlapped data (C), TGF-β1-treated DHBFCs (D), 1,25(OH)₂D₃-TGF-β1-treated DHBFCs (E), TGF-β1-treated DHBFCs vs 1,25(OH)₂D₃-TGF-β1-treated DHBFCs, overlapped data (F). The population percentage in each quadrant is also indicated. (G) Graphic quantitation of the respective cell populations using GraphPad. (H) mRNA expression of BAX in 1,25(OH)₂D₃ (50 nM) treated HBFCs. The Student’s t-test (variances assumed equally by ANOVA) was used to determine the difference between NHBFCs and DHBFCs under a given treatment and associated p- values are indicated. (ns) p > 0.05, no significant difference. Data were expressed as mean ± standard error (SE) from duplicate values of two independent experiments. NHBFCs (n=4), DHBFCs (n=4).