Fig. 4

Upregulating phospholipid synthesis or constraining precursor influx restores autophagy in TAG production defective cells. Autophagy was assessed by formation of GFP-Atg8 puncta (a, b), formation of Atg1 and Atg5 puncta (c, d), proteolytic processing of GFP-Atg8 (e), and pho8Δ60 assay (f–k). Induction of phospholipid production and reduction of precursor influx were achieved as in Fig. 3. a, c Representative microscopy images presented as in Fig. 1a. Cells starved for 1 h. b, d Quantification of Atg8, Atg1, and Atg5 puncta per cell in a, c. Error bar, standard deviation, n = 3. e Representative immunoblots from three independent repeats. Cells starved for 2 h. f–k pho8Δ60 enzymatic assay. Cells starved for 4 h. Error bar, standard deviation, n = 3