Mouse anti fbp2

The immunofluorescence studies were performed as described previously [18 (link)]. The cells were fixed, permeabilized, and incubated overnight at 4 °C with the following respective primary antibodies: rabbit anti-Fbp2 (1:500, produced and tested as described previously [19 (link)]), mouse anti-Fbp2 (1:500, Santa Cruz Biotechnology, Dallas, TX, USA, sc-271799), rabbit anti-Hif1α (1:500, Bioss Antibodies, Woburn, MA, USA, bs-0737R), rabbit anti-Ldha (1:500, Novus Biologials, Littleton, CO, USA, NBP1-48336), rabbit anti-Hk2 (1:500, Merck ab3279), rabbit anti-Ki67 (1:500, Abcam, Cambridge, UK, ab15580), mouse anti-αSMA (1:500, Merck a5228), mouse anti-β-actin (1:500, Sigma-Aldrich a1978), and rabbit anti-β-actin (1:500, Sigma–Aldrich a2066). The primary antibodies were detected using the following fluorophore-labelled secondary antibodies: goat anti-mouse-AlexaFluor488 (1:1000, Abcam, ab150113), goat anti-mouse-AlexaFluor633 (1:1000, Thermo Fisher Scientific, Waltham, MA, USA, a21050), goat anti-rabbit-AlexaFluor 488 (1:1000, Invitrogen a11034), and goat anti-rabbit-AlexaFluor633 (1:1000, Invitrogen a21070). Nuclei were counterstained with DAPI. In the controls, the primary antibodies were omitted.

Detection and visualization of Fbp-Camk2α interaction was performed using the DuoLink^® In Situ Orange Starter Kit Mouse/Rabbit (Merck KGaA, Darmstadt, Germany) according to the manufacturer’s instruction, using mouse anti-Fbp2 (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA, sc-271799) and rabbit anti-Camk2α (1:1000, Merck KGaA, Darmstadt, Germany, C6974) primary antibodies. In controls, the primary antibodies were omitted.