Cross-talk between microRNAs, nuclear factor E2-related factor 2, and heme oxygenase-1 in ochratoxin A-induced toxic effects in renal proximal tubular epithelial cells
Article first published online: 28 DEC 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Molecular Nutrition & Food Research
Volume 57, Issue 3, pages 504–515, March 2013
How to Cite
Stachurska, A., Ciesla, M., Kozakowska, M., Wolffram, S., Boesch-Saadatmandi, C., Rimbach, G., Jozkowicz, A., Dulak, J. and Loboda, A. (2013), Cross-talk between microRNAs, nuclear factor E2-related factor 2, and heme oxygenase-1 in ochratoxin A-induced toxic effects in renal proximal tubular epithelial cells. Mol. Nutr. Food Res., 57: 504–515. doi: 10.1002/mnfr.201200456
- Issue published online: 12 MAR 2013
- Article first published online: 28 DEC 2012
- Manuscript Accepted: 24 OCT 2012
- Manuscript Revised: 15 OCT 2012
- Manuscript Received: 12 JUL 2012
- Polish Ministry for Science and Higher Education. Grant Number: IP2011 031071
- National Science Center. Grant Numbers: N N401 297835, N N301 033440
- Foundation for Polish Science—Parent-Bridge Programme
- European Union within European Regional Development
- European Union and the Polish Ministry of Science and Higher Education. Grant Numbers: POIG.02.01.00–12-064/08 02.02.00–00-014/08, 01.01.02–00-069/09, 01.01.02–00-109/09
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Supporting Information Fig. S1. OTA down-regulates Nrf2 protein level in LLC-PK1 cells. LLC-PK1 cells were treated with 25 μM OTA for 24h and then immunofluorescent staining was performed.
Supporting Information Fig. S2. Adenoviral overexpression of Nrf2 and HO-1 does not influence cell morphology and does not cause cell death. LLC-PK1 cells were transduced with 50 MOI of AdNrf2 or AdHO-1 as wells as with AdGFP as a control vector. Increased mRNA (A) and protein (B, C) level of HO-1 and Nrf2 was observed without changes in morphology (D) and LDH release (E). A, mean of 4, E, mean of 3 experiments performed in duplicate, B - immunofluorescent staining, C - representative Western blots, D – representative photos, * p < 0.05 vs AdGFP-transduced cells
Supporting Information Fig. S3. DFO increases HIF activity and VEGF gene expression. LLC-PK1 cells were transfected with HRE-luc plasmid, and after 24h later stimulated with DFO (A). VEGF mRNA (B) and protein (C) level was assessed after 24h stimulation with 250 μM DFO by real-time PCR and ELISA, respectively. Mean of 3 experiments performed in duplicate
Supporting Information Fig. S4. OTA elevates expression of DGCR8 in LLC-PK1 cells. LLC-PK1 cells were treated for 24h with 25 μM OTA and then immunofluorescent staining was performed.
Supporting Information Fig. S5. Inhibition of miR-132 and miR-200c reduces OTA-mediated alterations in LLC-PK1 cells. LLC-PK1 cells were transfected with specific antagomirs (anti-miR-132 or anti-miR-200c) as well as with control anti-miR and after 24h were treated with 25 μM OTA for the next 24h. Inhibition of miR-132 counteracted the OTA-mediated reduction in Nrf2 protein (A). Immunofluorescent staining showed that both anti-miR-132 and anti-miR-200c increased the downregulated HO-1 protein level caused by OTA (B).
|mnfr1897-sup-0001-TableS1.doc||34K||Supporting Information Table S1. List of primers for gene expression analysis by real-time PCR|
|mnfr1897-sup-0002-TableS2.doc||34K||Supporting Information Table S2. List of primers for miRNAs expression analysis by real-time PCR|
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