Extended Data Fig. 1: Validation examples of CCD and non-CCD proteins using co-localization with mNG-tagged proteins and siRNA gene silencing.
From: Spatiotemporal dissection of the cell cycle with single-cell proteogenomics
a, The specificity of several dozen antibodies (45 in total with 10 examples presented here; Supplementary Table 3) targeting proteins with CCD expression was validated by co-localization with mNG-tagged protein. Scale bars, 10 μm. b, The specificity of several dozen antibodies (79 in total with 10 examples presented here; Supplementary Table 3) targeting proteins with non-CCD variable expression was validated by co-localization with mNG-tagged protein. Scale bars, 10 μm. c, The specificity of 29 antibodies (three presented here; Figs. 1c, 4b, Supplementary Table 3) was validated with siRNA-mediated gene silencing, which resulted in significantly lower staining intensity. Scale bars, 10 μm. For box plots: centre line, median; box, Q1 and Q3; whiskers, 1.5× IQR below Q1 and above Q3; points, outliers. TTC21B, P = 6.3 × 10−31; DUSP19, P = 0.0025; NET1, P = 9.7 × 10−8 by binomial one-sided tests. TTC21B, n = 620, 358; DUSP19, n = 125, 147; NET1, n = 162,1 49 independent cells for control and siRNA, respectively. d, The specificity of 321 antibodies was validated by independent antibody staining, as exemplified by SCIN. Scale bars, 10 μm.
