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. 2021 Jul 16;12:4357. doi: 10.1038/s41467-021-24574-y

Fig. 6. VUN103 and Nb7 have opposing effect on constitutive and ligand-induced signaling.

Fig. 6

a Effect of non-US28 targeting FLAG-tagged intrabody (Irr Nb), FLAG-tagged VUN103 (VUN103), and/or Nb7-mVenus (Nb7) on US28-mediated accumulation of inositol phosphate (IP) in US28-expressing HEK293T cells that were untreated or stimulated with 30 nM CX3CL1 (CX3CL1). Expression of FLAG-tagged intrabodies was induced using 100 nM of tebufenozide, one day prior to read-out. Cells expressing a single intrabody (Single intrabody exp) or two intrabodies (Double intrabody exp) are divided by a dashed line. Accumulation of inositol phosphates were normalized to untreated US28-expressing cells (n = 3 independent experiments). b Determination of Nb7-mVenus expression levels by measuring the fluorescence intensity of mVenus. Cells expressing a single intrabody (Single intrabody exp) or two intrabodies (Double intrabody exp) are divided by a dashed line (n = 3 independent experiments). c Expression of FLAG-tagged intrabody was determined via ELISA by the C-terminal FLAG-tag and an anti-FLAG antibody. Cells expressing a single intrabody (Single intrabody exp) or two intrabodies (Double intrabody exp) are divided by a dashed line (n = three independent experiments). All data are plotted as mean ± SD. Statistical analyses were performed using an unpaired two-tailed t-test. ns, p > 0.05. Source data are provided as a Source Data file.