ULK1 and infection: Infection of atg1- cells with either M. marinum ΔRD1 or ΔCE, a strain with a knock out restricted to ESAT-6 and its chaperone CFP-10 [41], also showed reduced bacteria ubiquitination at 6–7 hpi (Figs 4C and S4D), suggesting that secretion of ESAT-6 is thus required for cytosolic escape and subsequent ubiquitination of mycobacteria in D. discoideum. Complementation of the ΔRD1 strain with the M. tuberculosis-derived RD1-2F9 cosmid [42] restored ubiquitination to wt levels (Fig 4C).