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. 2020 Mar 3;10(3):393. doi: 10.3390/biom10030393

Figure 2.

Figure 2

(A) Multiplex assay for human monocyte cell line U937 treated with and without Hcy 20 μM for 24 h revealed significant increase in pro-inflammatory cytokines (IP-10, TNFα, IL-B, and IL-6). (B–E). HHcy activates NF-κB transcription factor in cultured HRECs and ARPE-19 cells. Western blot analysis for HRECs (B,D) and ARPE-19 cells (C,E) treated with 100 µM Hcy, showing significant increase in the NF-κB nuclear to cytoplasmic ratio compared to the control untreated cells. NF-κB is a transcription factor, which controls the transcription of DNA and cytokine production. (F) IF staining for NF-κB, showed that Hcy activated and translocated NF-κB from the cytoplasmic to the nuclear side in HRECs treated with Hcy (green). (G) Primary RPE cells isolated from cbs+/− mice retina (red). (H–I) ELISA measurement of NFκB (P65) in HRECs (H) and ARPE-19 (I) treated with and without Hcy (50 μM and 100 μM) for 24 h. Samples were representative to at least three mice for each experiment. Data are expressed as mean ± SD. (* p < 0.05, ** p < 0.01). Scale bar: 50 and 20 μm, respectively.