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MEFs were grown on coverslips and fixed with methanol for 2 min. After two PBS washes, MEFs were saturated with the blocking solution followed by a 1 h at 37 °C incubation with the primary antibodies: Bcl-xL (Cell Signaling #2764, 1/1000) and IP3R C-terminal (abcam #ab190239, 1/1000). PLA secondary antibodies probes were added and incubated for 1 h at 37 °C after 3 PBS washes. Ligation and amplification (100 min at 37 °C) steps followed. Cells were mounted with Duolink II Mounting medium containing DAPI and images were acquired using the fluorescence microscope.
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