Figure S4.
Effects of UBQLN single KD on the CFTR ERAD, triple KD on CFTR retrotranslocation, and establishment of TCRα-HiBiT and Insig-1-HiBiT ERAD assay. (A) The kinetic degradation of ∆F508-CFTR-HiBiT(CT) in 293MSR WT cells transfected with 50 nM siNT or siUNQLN1, 2, or 4. The ERAD rate was calculated by fitting the initial degradation portion of each kinetic degradation curve (right, n = 2). Each biological replicate (n) is color-coded: the averages from four technical replicates are shown in triangles. Data represent mean. (B) The retrotranslocation of ∆F508-CFTR-HiBiT(Ex) in 293MSR cells upon UBQLN1/2/4 triple KD was measured during the MG-132 and CHX chase (n = 3, unpaired t test). (C and D) The HiBiT degradation assay confirmed the proteasomal degradation of TCRα-HiBiT (C, n = 4) and Insig-1-HiBiT (D, n = 3) in 293MSR cells. Data represent mean ± SD. **P < 0.01.

Effects of UBQLN single KD on the CFTR ERAD, triple KD on CFTR retrotranslocation, and establishment of TCRα-HiBiT and Insig-1-HiBiT ERAD assay. (A) The kinetic degradation of ∆F508-CFTR-HiBiT(CT) in 293MSR WT cells transfected with 50 nM siNT or siUNQLN1, 2, or 4. The ERAD rate was calculated by fitting the initial degradation portion of each kinetic degradation curve (right, n = 2). Each biological replicate (n) is color-coded: the averages from four technical replicates are shown in triangles. Data represent mean. (B) The retrotranslocation of ∆F508-CFTR-HiBiT(Ex) in 293MSR cells upon UBQLN1/2/4 triple KD was measured during the MG-132 and CHX chase (n = 3, unpaired t test). (C and D) The HiBiT degradation assay confirmed the proteasomal degradation of TCRα-HiBiT (C, n = 4) and Insig-1-HiBiT (D, n = 3) in 293MSR cells. Data represent mean ± SD. **P < 0.01.

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