Fig 1: RELB affects proliferation, apoptosis, and migration in hRECs. (A) CCK-8 analysis showing that RELB knockdown inhibited the proliferation of hRECs. (B) FCM analysis showing that after transfection with siRELB, apoptosis and cell death significantly increased in hRECs. (C) As shown by the Transwell assay, the cells were stained with crystal violet; the knockdown of RELB repressed the migration of hRECs, the scale bar was 500 µm. (D,E) Transfection with siRELB reduced the wound healing process, the scale bar was 200 µm. (F,G) siRELB decreased the tube formation process, the scale bar was 200 µm. Error bars represent the mean ± SD of at least triplicate experiments. *, P<0.05. (H) Western blotting analysis of protein levels of RELB, p100, p52, Bax, and Bcl-2. After transfection with siRELB, the protein level of RELB was decreased, whereas that of p100 was increased and p52 was decreased; for proteins in the canonical apoptosis pathway, Bcl-2 was decreased and the level of Bax was increased. RELB, reticuloendotheliosis viral oncogene related B; hRECs, human retinal microvascular endothelial cells; CCK-8, Cell Counting Kit-8; FCM, flow cytometry; HG, high glucose; LG, low glucose.
Fig 2: Rescue effects of LTBR on si-CREB1 in the viability, apoptosis, and NF-?B pathway in hyperoxia-stimulated ATII cells. (a) Cell viability of hyperoxia-treated A549 and ATII cells after transfection of si-NC, si-CREB1, and cotransfection of si-CREB1 + pcDNA-LTBR. (b, c) Flow cytometry results in A549 and ATII cells after indicated transfections. (d) Quantification analysis of the flow cytometry results in b and c to show cell apoptosis rate. (e) The ratio of p-IKKa/IKKa and relative grey density of p52 and RELB in hyperoxia-stimulated A549 and ATII cells were measured by western blotting. ***p < 0.001.
Fig 3: Si-LTBR inactivates the NF-?B pathway. (a, b) Protein levels of p-IKKa, IKKa, p52, and RELB in A549 and ATII cells were detected by western blotting. The ratio of p-IKKa/IKKa and relative grey density of p52 and RELB were shown in the graph bars following the protein bands. **p < 0.01, ***p < 0.001.
Fig 4: Si-LTBR rescues the effects of pcDNA-CREB1 in the viability, apoptosis, and NF-?B pathway-associated proteins in hyperoxia-stimulated ATII cells. (a) Cell viability of hyperoxia-treated A549 and ATII cells after transfection of pcDNA-NC, pcDNA-CREB1, and cotransfection of pcDNA-CREB1 + si-LTBR. (b, c) Flow cytometry results in A549 and ATII cells after indicated transfections. (d) Quantification analysis of the flow cytometry results in b and c to show cell apoptosis rate. (e) The ratio of p-IKKa/IKKa and relative grey density of p52 and RELB in hyperoxia-stimulated A549 and ATII cells were measured by western blotting. **p < 0.01, ***p < 0.001.
Supplier Page from Abcam for Anti-NFkB p100/NFKB2 antibody [EPR4686-66]