1. Additional file 1 of TRIM46 aggravated high glucose-induced hyper permeability and inflammatory response in human retinal capillary endothelial cells by promoting IκBα ubiquitination
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Shen, Hangqi, Gong, Qiaoyun, Zhang, Jingting, Wang, Haiyan, Qiu, Qinghua, Zhang, Jingfa, and Luo, Dawei
- Abstract
Additional file 1: Fig. S1 TRIM46 overexpression and short hairpin RNA (shRNA)-mediated interference of TRIM46 in HRCECs. a Protein expression of TRIM46 in HRCECs transfected with oeTRIM46 or Vector. b Protein expression of TRIM46 in HRCECs transduced with TRIM46 shRNAs (shTRIM46-1, shTRIM46-2 and shTRIM46-3) or control shRNA (shNC). HRCECs, human retinal capillary endothelial cells; TRIM, tripartite motif. Fig. S2. The images of the monolayers are shown (scale bar: 25 μm). a In HRCECs, TRIM46 was interfered with or overexpressed for 24 h. After the cells grew to confluence, the cells were then treated with HG (25 mM). Control osmotic pressure was controlled with normal glucose concentration of 5.5 mM and mannitol. b HRCECs transfected with oeTRIM46 or Vector with overexpression of IκBα (oeIκBα). After the cells grew to confluence, cell permeability analysis was performed (Figure 3d-e). c HRCECs were transfected with oeTRIM46 or Vector. After the cells grew to confluence, cells were administered with the NF-κB inhibitor PDTC (10 μM) and HG (25 mM). Cell permeability analysis was then performed (Figure 4e-f). HG, high glucose; HRCECs, human retinal capillary endothelial cells; NF-κB, nuclear factor kappa B; PDTC, pyrrolidine dithiocarbamate; TRIM, tripartite motif. Fig. S3. Immunofluorescence staining of tight junction proteins ZO-1 and Occludin. Blue: DAPI, Green: ZO-1 or Occludin. Scale bar: 50 μm. a In HRCECs, TRIM46 was interfered with or overexpressed for 24 h. After the cells grew to confluence, the cells were then treated with HG (25 mM). Control osmotic pressure was controlled with normal glucose concentration of 5.5 mM and mannitol. b HRCECs transfected with oeTRIM46 or Vector with overexpression of IκBα (oeIκBα). After the cells grew to confluence, cell permeability analysis was performed (Figure 3d-e). c HRCECs were transfected with oeTRIM46 or Vector. After the cells grew to confluence, cells were administered with the NF-κB inhibitor PDTC (10 μM) and HG (25 mM). Cell permeability analysis was then performed (Figure 4e-f). HG, high glucose; HRCECs, human retinal capillary endothelial cells; NF-κB, nuclear factor kappa B; PDTC, pyrrolidine dithiocarbamate; TRIM, tripartite motif.
- Published
- 2022
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