FIGURE 4:

Endogenous ERManI is a Golgi-resident protein. (A) Confocal images of HeLa cells coimmunostained with the anti-ERManI mAbs and anti-Giantin or anti-calnexin antibodies. ERManI was visualized using Alexa 488–conjugated anti–mouse secondary antibody (green). Giantin or calnexin was visualized using Alexa 555–conjugated anti–rabbit secondary antibodies (red). Cell nuclei were counterstained with To-PRO3. (B) Flag-tagged ERManI was transfected into HeLa cells. Forty-eight hours after transfection, cells were fixed and coimmunostained with anti-Flag and anti-Giantin antibodies. Flag-ERManI was visualized using Alexa 488–conjugated anti–mouse secondary antibody (green). Giantin was visualized using Alexa 555–conjugated anti–rabbit secondary antibodies (red). Arrows indicate the cells transfected with Flag-ERManI. (C) HeLa cells treated with 10 μM MG132 for 6 h were costained for ERManI (green) and BAP31 (red). Cell nuclei were counterstained with DAPI.