Supplementary Figure 2: SCRIB is a component of the NOX complex. | Nature Cell Biology

Supplementary Figure 2: SCRIB is a component of the NOX complex.

From: An interaction between Scribble and the NADPH oxidase complex controls M1 macrophage polarization and function

Supplementary Figure 2

(a) Quantitation of SCRIB binding to β-PIX. (b) Wild type (WT), Flagp22phox (Flag-p22) or Flag-p22phox without last four amino acid (Flag-p22Δ) were co-transfected with T7 tagged full length SCRIB. Flag immunoprecipitants were recovered and analyzed by anti-Flag and anti-T7 immunoblots. n = 3 biologically independent experiments (mean ± SD). (two-tailed, unpaired t-test with equal variances: P = 0.001). (c) Components of the four NOX complexes, NOX1-4. (d) Total cell lysates (TCL) from 293T cells transfected with Myc-tagged NOX complex proteins or anti-Myc immunoprecipitates were analyzed for the presence of endogenous SCRIB. (e) Total cell lysates (TCL) from 293T cells transfected with Myc-tagged NOX complex proteins or anti-Myc immunoprecipitates were analyzed for the presence of Myc-tagged proteins. (f) SCRIB immunoprecipitation followed by anti p22phox immunoblots using lysates obtained from parental RAW 264.7 cell line. Quantitation of p22phox binding to SCRIB in RAW 264.7 cell line (g) or in bone marrow derived macrophages (h) stimulated with PMA. Each experiment was performed in triplicate and data presented is an average of n = 3 independent experiments (mean ± SD). Unprocessed scans of full blots/gels are shown in Supplementary Fig. 6.

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