Crosstalk between the Unfolded Protein Response and NF-κB-Mediated Inflammation in the Progression of Chronic Kidney Disease

Figure 2

Detection of XBP1 splicing in human proximal tubule-2 cells using XBP1sVenus reporter. (a) XBP1sVenus was placed in a hygromycin B selectable cassette and transfected into HK-2 cells. Cells were then selected for stable incorporation of XBP1sVenus with 0.5 mg/mL hygromycin B. Clones were screened with 1 μg/mL tunicamycin (Tm) for 6 or 18 hours. Clones 3, 12, and 16 were found to give robust responses when probed with the flag-tag antibody and produced green fluorescence. (b) Clone 12 from the stably transfected XBP1s HK-2 reported cell line was grown up and untreated (Un) or treated with 1 μg/mL tunicamycin (Tm). The expression of the flag-tag shows IRE1 activation in the reporter cell line.

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