ITC replicates and co-IP with WT and 3-Mut Ig 1–2. (A) Isothermal titration calorimetry (ITC) replicates from Fig. 3 between WT Ig 1–2 and Ig 10–11. (B) ITC replicates for 3-Mut Ig 1–2 and Ig 10–11. Dissociation constants (KD) are indicated. n = 3 independent replicates were performed for each condition. (C) Immunoblot for expression of WT Ig 1–2-HA, 3-Mut Ig 1–2-HA, and Ig 10–11-FLAG in U2OS cells used for quantification in Fig. 3 G. (D) Co-immunoprecipitation (co-IP) analysis of interactions between HA-tagged Ig 1–2 constructs and WT or 4-Mut FL NOMO. Whole-cell lysates (input) and HA immunoprecipitates (IP: HA) were analyzed by immunoblot using antibodies against FLAG, NOMO, and HA. GAPDH serves as a loading control for input samples. Unprocessed blots are available in source data. Source data are available for this figure: SourceData FS3.
Figure S3.

ITC replicates and co-IP with WT and 3-Mut Ig 1–2. (A) Isothermal titration calorimetry (ITC) replicates from Fig. 3 between WT Ig 1–2 and Ig 10–11. (B) ITC replicates for 3-Mut Ig 1–2 and Ig 10–11. Dissociation constants (KD) are indicated. n = 3 independent replicates were performed for each condition. (C) Immunoblot for expression of WT Ig 1–2-HA, 3-Mut Ig 1–2-HA, and Ig 10–11-FLAG in U2OS cells used for quantification in Fig. 3 G. (D) Co-immunoprecipitation (co-IP) analysis of interactions between HA-tagged Ig 1–2 constructs and WT or 4-Mut FL NOMO. Whole-cell lysates (input) and HA immunoprecipitates (IP: HA) were analyzed by immunoblot using antibodies against FLAG, NOMO, and HA. GAPDH serves as a loading control for input samples. Unprocessed blots are available in source data. Source data are available for this figure: SourceData FS3.

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