Fig. 8

In situ detection of miR34c in senescent cells. Detection of miR34c in senescent cells employing the HBEC CDC6 Tet-ON system. Double staining was performed in two consecutive states: in the OFF state where proliferation of HBECs is evident (“OFF”) and 6 days after constitutive induction of CDC6 when cells are senescent (“6d ON”). Step 1: Fluorescence FISH of miR34c employing a double-DIG-labeled LNA probe, visualized as green emission in the cytoplasm, using TSA plus Fluorescein (emitting at 518 nm). Step 2: GL13 staining, visualized at far red spectra as granules in the cytoplasm employing Alexa Fluor goat-anti-mouse (647 nm) (emitting at 668 nm). Scale bar: 50 μm