JCS -- Goetz et al. 120 (20): 3553 Figure IG2

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Fig. 2. Characterisation of the 3F3A-labelled ER domain. (A) Cos-7 cells were co-transfected with GFP-Sec61 $beta$ and Rtn4a-MYC and visualised using the GFP tag and anti-MYC and 3F3A antibodies. Inset shows that 3F3A labelling does not overlap with the Rtn4a-labelled ER tubules or the perinuclear ER labelled for GFP-Sec61 $beta$ . (B) Cos-7 cells transfected with Rtn4a-MYC were labelled with anti-MYC, 3F3A and anti-Sec61 ${alpha}$ antibodies. Rtn4a distribution is distinct from both 3F3A and Sec61 ${alpha}$ , and 3F3A shows only partial colocalisation with Sec61 ${alpha}$ . Alternatively, FLAG-AMFR-transfected cells were immunofluorescently labelled for FLAG, 3F3A and either Sec61 ${alpha}$ (C) or mtHSP70 (E) or co-transfected with vector encoding Rtn4a-MYC and labelled for FLAG, 3F3A and MYC (D). Insets highlight colocalisation of 3F3A labelling with anti-FLAG (C), Rtn4a (D) and mtHSP70 (E). Cells labelled for Sec61 ${alpha}$ were pre-treated with RNAse. Bars, 20 µm; inset bars, 5 µm.