Anti mcherry antibody

Semi-synchronous predation experiments were set up as above. Samples of 1.4 ml were pre-fixed in 0.25% paraformaldehyde and recovered by centrifugation at 17,000 × g for 2 min, then fixed in 2.5% paraformaldehyde in Dulbecco’s PBS for 10 min at 37 °C. Cells were recovered by centrifugation at 17,000 × g for 2 min, washed twice in 100 µl blocking solution (2% bovine serum albumin, BSA, in Dulbecco’s phosphate buffered saline, PBS) and then incubated in blocking solution for 45 min. Samples were further incubated in anti-mCherry antibody (Invitrogen PA5-34974) diluted 1:1,000 in blocking solution for 1 h. Cells were recovered by centrifugation at 17,000 × g for 2 min, then washed twice in blocking solution before a final incubation with secondary antibody of goat anti-rabbit IgG with Alexa Fluor plus 488 (Invitrogen A32731). Cells were then washed twice with blocking solution before imaging with a Nikon Ti-E microscope as described above. Some samples were further stained with FM46-4 by resuspension in 10 mM stain in water. Images were analysed using ImageJ software (Fiji). Adjustments to brightness and contrast for whole images were made until FM46-4-stained flagella and anti-mCherry foci were visible. Cells were then manually (visually) scored for the presence of flagella (FM46-4 red channel) and the presence and positioning of anti-mCherry foci (GFP channel).

The anti-LmrC(2) antibody was generated by GenScript USA, Inc., by inoculating a New Zealand rabbit host strain with a peptide-KLH conjugate containing the LmrC peptide CLQRQAQESAGRAAS. The specificity of the LmrC antibody was validated using S. lincolnensis WT and ΔC mycelium from 16-h seed cultures (see Fig. S7c) grown in the absence or presence of lincomycin (LIN; 4 mg liter⁻¹) or clindamycin (CLI; 0.5 mg liter⁻¹) (see Fig. S5a). The anti-m-Cherry antibody was purchased from Invitrogen (Cat PA5-34974) and AB Clonal. The ribosomal protein S7-specific antibody was obtained from Mee-Ngan F. Yap (72 (link)).

Anti-TRIM44 polyclonal antibody (Proteintech Group, 11511–1-AP); anti-Ub antibody (Biolegend, 646301); anti-mCherry antibody (ThermoFisher, PA5–34974); anti-NFE2L2 antibody (ThermoFisher, PA5–27882); anti-ATG5 antibody (Novus Biologicals, NBP2–24389); anti-GFP antibody (Santa Cruz Biotechnology, sc-9996); anti-ACTB antibody (Santa Cruz Biotechnology, sc-47778), anti-HA antibody (Santa Cruz Biotechnology, sc-805); anti-LC3B antibody (Cell Signaling Technology, 3868); anti-SQSTM1 antibody (Cell Signaling Technology, 88588); anti-LaminB1 antibody (R &D, MAB8525); anti-KEAP1 antibody (Origene, TA502059); anti-GAPDH antibody (Invitrogen, PA5–85074); anti-FLAG antibody (Invitrogen, PA1–984); anti-mTOR antibody (CST, 2972); anti-phospho-mTOR (Ser2448) antibody (Millipore Sigma, 09–213) anti-SQSTM1 antibody(CST, 7695,88588); anti-phospho-SQSTM1/p62 (Ser349) (E7M1A) antibody (CST, 16177S); anti-phospho-SQSTM1/p62 (Ser403) (D8D6T) antibody (CST, 39786S); anti-PKA Cα antibody (CST, 4782); ; anti-PKA substrate antibody (CST, 9624); Goat anti-Mouse IgG (Invitrogen, 31430); Goat anti-Rabbit IgG (Invitrogen, 31460); Alexa Fluor 594 donkey anti rabbit IgG (H + L) (Invitrogen, A21207); Alexa Fluor 546 goat anti mouse IgG (H + L) (Invitrogen, A11003).