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. Author manuscript; available in PMC: 2020 Jul 21.
Published in final edited form as: Cell Rep. 2020 Jun 30;31(13):107840. doi: 10.1016/j.celrep.2020.107840

Figure 4. The N-Glycan Content of GRP94 Regulates the Stability and Function of the HMW GRP94-HER2 PM-Associated Pool.

Figure 4.

(A) Schematic of the experimental design to investigate the role of N-glycans on the conformation, stability, and function of the HMW GRP94 pools. WS13-B, biotinylated PU-WS13.

(B) PM pool of GRP94 and HER2 in SKBr3 extracts treated under native conditions as in (A) with the indicated enzymes. Graph, mean; error bars, SEM; n = 5; unpaired t test, ***p < 0.001, ****p < 0.0001.

(C) Western blot analysis of GRP94 and HER2 isolated from extracts treated as in (A) and (B). Graph, mean; error bars, SEM; n = 3 to 5; unpaired t test, **p < 0.01, *p < 0.05.

(D) Western blot analysis of cells treated for 4 h with PU-WS13 (10 μM) prior to N-glycan removal and immuno-capture with the G4420 anti-GRP94 antibody as in (A). Graph, mean; error bars, SEM; n = 3; unpaired t test, **p < 0.01.

(E) Biochemical signature of HER2 in extracts as in (C) and (D).

(F) Summary of the findings.

See also Figure S4.