Supplementary Figure 2: Quantification of tubulin detyrosination assay immunoblots. | Nature Structural & Molecular Biology

Supplementary Figure 2: Quantification of tubulin detyrosination assay immunoblots.

From: Structural basis of tubulin detyrosination by the vasohibin–SVBP enzyme complex

Supplementary Figure 2

Quantification results of immunoblots corresponding to Supplementary Fig. 1c (a) and Figs 2c (b), 2f (c), 3c (d), 4f (e), 4g (f), and 5b (g). Immunoblots, resulting each from an independent transfection, were analyzed for the negative control (sham transfection, white column, n = 16 total), the wild type non-mutated vasohibins (black columns, n = 3–5 per graph) or mutated vasohibins (gray columns, n = 3 except for F186A and Y236A in panel a (n = 2), and W63E W67E in panel e (n = 4)), and expressed as mean ± sem. In the top graphs, the plotted values represent the relative intensities of the detyrosinated-tubulin (deTyr-tub) signal obtained with mutants after normalization to the GAPDH signal. The signal of the wild type vasohibin from the same blots was set to 1.0, upper dotted line. Note that this measurement corresponds to endogenous (bottom dotted line) plus exogenous detyrosination activities in cells. In the middle graphs, the plotted values represent the relative quantities of mutant vasohibins expressed in cells after normalization to the GAPDH signal. The signal of the wild type vasohibin from the same blots was set to 1.0, upper dotted line. Note that this signal only corresponds to exogenous vasohibin levels since detection was performed using an anti-eGFP antibody. In the bottom graphs, the plotted values represent the relative intensities of the deTyr-tubulin signal obtained with mutants after subtracting the HEK293T cells endogenous levels of deTyr-tubulin (obtained with sham transfected cells, white columns in top graphs). The signal obtained with wild type vasohibin from the same blots was set to 1.0, upper dotted line. All The source files are available in Supplementary Data Set 2.

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