Fig. 6: In vitro cell uptake and retention of A947.
From: Tissue distribution and retention drives efficacy of rapidly clearing VHL-based PROTACs
a Time-dependent uptake of A947 in hepatocytes in the absence and presence of pan-solute carrier transporter (SLC) inhibitor rifamycin SV (n = 3) (solid blue circle, 37 °C A947; solid red square, 37 °C A947+rifamycin; solid pink triangle, 4 °C A947). The cellular concentration was measured by LC/MS and represented as peak area ratios to the internal standard. Data are normalized to DMSO control treated cultures and are presented as mean ± SD from three independent replicates. Individual numerical data are listed in Supplementary Table 14). b Evaluation of re-synthesis of SMARCA2 protein in NCI-H1944 cells following washout of A947 (n = 5). Quantification of SMARCA2 protein levels in cell by immunofluorescence following treatment and washout of A947. Data are normalized to DMSO control treated cultures and are presented as mean ± SD from five independent replicates. The VHL ligand, A2702, was spiked in at 1000× molar excess following washout to compete with residual A947 in cells. SMARCA2 levels were evaluated at 48 h and 168 h following washout.
