Journal: bioRxiv

Article Title: Urokinase receptor associates with TLR4 interactome to promote LPS response

doi: 10.1101/2020.06.10.143826

Figure Lengend Snippet: A. Downregulation of uPAR expression in mouse mesothelial epithelial cells. B. LPS response of SiCo and uPARsi mouse mesothelial cells was assessed after stimulation with 100ng/ml LPS for 3 hrs. Expression was analysed by TaqMan RT-PCR. C. LPS-induced protein tyrosine phosphorylation was assessed in SiCo and uPARsi mouse mesothelial cells by western blotting of the whole cell lysate with P-Tyr antibody (upper panel) and P-p65 antibody (middle panel). GAPDH shows loading control (lower panel). D. Quantification of tyrosine (left) and p65 phosphorylation (right) from three independent western blotting experiments.

Article Snippet: HK-2 human kidney proximal tubule epithelial cells were from ATCC and cultivated as recommended by the supplier in Keratinocyte Serum Free Medium containing 0.05 mg/ml bovine pituitary extract and 5 ng/ml EGF.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Control

Journal: bioRxiv

Article Title: Urokinase receptor associates with TLR4 interactome to promote LPS response

doi: 10.1101/2020.06.10.143826

Figure Lengend Snippet: A, B. LPS-induced expression of IL-6 and IL-8 by human renal proximal tubule epithelial cell (HK-2) was assessed by TaqMan RT-PCR (A and ELISA (B). C. HMGB1-dependent IL-6 and IL-8 expression by HK-2 cells was assessed by TaqMan RT-PCR. D. Human renal epithelial HK-2 cells were lentivirus-infected to express Gaussia luciferase under control of NFκB and GAPDH promoters. Enzyme activity was measured in cell conditioned media 10 hrs after stimulation with LPS.

Article Snippet: HK-2 human kidney proximal tubule epithelial cells were from ATCC and cultivated as recommended by the supplier in Keratinocyte Serum Free Medium containing 0.05 mg/ml bovine pituitary extract and 5 ng/ml EGF.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Infection, Luciferase, Control, Activity Assay

Journal: Acta Pharmacologica Sinica

Article Title: Involvement of endoplasmic reticulum stress in angiotensin II-induced NLRP3 inflammasome activation in human renal proximal tubular cells in vitro

doi: 10.1038/aps.2015.21

Figure Lengend Snippet: Angiotensin II induced inflammasome activation in tubular epithelial cells. (A) Real-time PCR and (C, E, and G) western blot analysis show that the mRNA and protein expression of caspase-1, IL-1β, and IL-18 were increased after treatment with different amounts (0, 10, 100, or 1000 nmol/L) of angiotensin II for 12 h in serum-free medium. (B) Real-time PCR and (D, F, and H) western blot analysis indicate that the mRNA and protein expression of caspase-1, IL-1β, and IL-18 were increased after treatment with 100 nmol/L angiotensin II for various time periods (0, 6, 12, or 24 h) in serum-free medium. β-Actin served as an internal control gene. The results represent the mean±SD from four experiments. bP<0.05 vs control.

Article Snippet: Human kidney proximal epithelial cells (HK-2) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Control