Fig. 6: Localized PKA RII is responsible for the effects of cAMP on autophagy.

A HT-29 cells were challenged with FSK (5 µM) or FSK (20 µM) combined to IBMX (500 µM) and total cell lysates were prepared at different timepoints. The phosphorylation status of endogenous PKA substrates was assessed by a phospho-PKA substrate-specific antibody, RRX(S/T)^P and is summarized in the inset (bar graphs: average ± SD of three independent experiments). B Summary of the effects on YFP-LC3 puncta of FSK 5 µM or FSK 20 µM combined to IBMX (500 µM) treatments, normalized to vehicle control (DMSO), measured after 4–6 h after treatment. Average of at least three independent experiments. C Summary of the effects of super AKAP-IS-mCherry (sAKAP-IS), RIAD-mCherry (RIAD), and scramble-mCherry (scr) on the number of YFP-LC3 puncta generated by FSK (20 µM)/IBMX (500 µM). Data presented are average of four independent experiments for sAKAP-IS, RIAD, and scramble, and ten experiments for control (15–25 cells for each treatment).