Figures & data
Table 1. Primers used in this study.
Figure 1. Tofacitinib suppresses ox-LDL-induced attachment of human monocyte THP-1 cells to HAECs. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. Representative fluorescence microscopic images of THP-1 cells attached to HAECs and quantitative analysis. Scale bar, 50 μM. (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).
![Figure 1. Tofacitinib suppresses ox-LDL-induced attachment of human monocyte THP-1 cells to HAECs. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. Representative fluorescence microscopic images of THP-1 cells attached to HAECs and quantitative analysis. Scale bar, 50 μM. (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).](/cms/asset/17a87241-58b3-4ed2-befc-4929077ff880/ianb_a_1573740_f0001_c.jpg)
Figure 2. Tofacitinib inhibits ox-LDL-induced expression of VCAM-1 and E-selectin. (A) Real-time PCR analysis of VCAM-1 and E-selectin; (B) Western blot analysis of VCAM-1 and E-selectin (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6). HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h.
![Figure 2. Tofacitinib inhibits ox-LDL-induced expression of VCAM-1 and E-selectin. (A) Real-time PCR analysis of VCAM-1 and E-selectin; (B) Western blot analysis of VCAM-1 and E-selectin (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6). HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h.](/cms/asset/b975a56e-eba0-4de5-b2ad-d7917078da0a/ianb_a_1573740_f0002_b.jpg)
Figure 3. Tofacitinib decreases ox-LDL-induced expression of LOX-1. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A) Real-time PCR analysis of LOX-1; (B) Western blot analysis of LOX-1 (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).
![Figure 3. Tofacitinib decreases ox-LDL-induced expression of LOX-1. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A) Real-time PCR analysis of LOX-1; (B) Western blot analysis of LOX-1 (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).](/cms/asset/0a65b558-4b7f-4668-bc53-8206b575cd11/ianb_a_1573740_f0003_b.jpg)
Figure 4. Tofacitinib suppresses ox-LDL-induced production of the pro-inflammatory cytokines TNF-α and IL-1β. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A) Real-time PCR analysis of TNF-α and IL-1β; (B). ELISA analysis of TNF-α and IL-1β (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).
![Figure 4. Tofacitinib suppresses ox-LDL-induced production of the pro-inflammatory cytokines TNF-α and IL-1β. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A) Real-time PCR analysis of TNF-α and IL-1β; (B). ELISA analysis of TNF-α and IL-1β (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).](/cms/asset/b8ce6cbe-a688-4631-96ef-e60a50599c61/ianb_a_1573740_f0004_b.jpg)
Figure 5. Tofacitinib ameliorates ox-LDL-induced generation of reactive oxygen species (ROS). HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. Intracellular ROS was determined by the DCFH-DA assay. Scale bar, 100 μM. (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM Tofacitinib group, ANOVA, n = 5–6).
![Figure 5. Tofacitinib ameliorates ox-LDL-induced generation of reactive oxygen species (ROS). HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. Intracellular ROS was determined by the DCFH-DA assay. Scale bar, 100 μM. (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM Tofacitinib group, ANOVA, n = 5–6).](/cms/asset/1864562e-8a5c-4ee8-9b22-6ac114eb7b1d/ianb_a_1573740_f0005_c.jpg)
Figure 6. Tofacitinib ameliorates ox-LDL-induced LDH release and reduction of cell viability. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 48 h. (A). LDH release was determined using a commercial kit; (B) Cell viability was determined by MTT assay (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).
![Figure 6. Tofacitinib ameliorates ox-LDL-induced LDH release and reduction of cell viability. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 48 h. (A). LDH release was determined using a commercial kit; (B) Cell viability was determined by MTT assay (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, $, p < .01 vs. ox-LDL + 100 nM tofacitinib group, ANOVA, n = 5–6).](/cms/asset/e96d7f21-034b-45a8-bb19-f8d10641143f/ianb_a_1573740_f0006_b.jpg)
Figure 7. Tofacitinib mitigates ox-LDL-induced phosphorylation and degradation of IκBα. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (500 nM) for 6 h. Phosphorylated and total levels of IκBα were determined by Western blot analysis (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, ANOVA, n = 5–6).
![Figure 7. Tofacitinib mitigates ox-LDL-induced phosphorylation and degradation of IκBα. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (500 nM) for 6 h. Phosphorylated and total levels of IκBα were determined by Western blot analysis (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, ANOVA, n = 5–6).](/cms/asset/66bcc539-b00a-4439-827f-15a69faa2426/ianb_a_1573740_f0007_b.jpg)
Figure 8. Tofacitinib inhibits ox-LDL-induced activation of NF-κB. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A). Nuclear level of p65; (B). Luciferase activity of NF-κB promoter (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, ANOVA, n = 5–6).
![Figure 8. Tofacitinib inhibits ox-LDL-induced activation of NF-κB. HAECs were stimulated with 100 mg/L ox-LDL in the presence or absence of tofacitinib (100, 500 nM) for 24 h. (A). Nuclear level of p65; (B). Luciferase activity of NF-κB promoter (*, p < .01 vs. vehicle control; #, p < .01 vs. ox-LDL group, ANOVA, n = 5–6).](/cms/asset/8478ccd4-9ea0-4bd9-83d9-2ea008803210/ianb_a_1573740_f0008_b.jpg)