Fig. 4

Phloretin stimulates autophagy in an AMPK-dependent manner. A Downstream analysis of the RNA-seq data obtained from phloretin treated BMDMs stimulated with LPS illustrated autophagy as one of the downstream biological processes that is activated by phloretin (z-score: 0.779). Data are represented by a heat map containing the normalized counts of genes associated with autophagy. A colour gradient was used to indicate the normalized counts and corresponding Fold change (Fc) differences, per sample and gene respectively. B–D Quantification and representative images of LC3 and p62 staining in phloretin-treated BMDMs stimulated with bafilomycin A1 (n = 5). The dotted line represents untreated cells (controls). E, F Western blot quantification and representative bands of the autophagy markers LC3II and p62 in phloretin-treated BMDMs stimulated with bafilomycin A1 alone or bafilomycin and the AMPK inhibitor together (n = 2). The dotted line represents cells treated only with bafilomycin A1 (controls). Ctrl, control; phl, phloretin; baf, bafilomycin A1. Data are represented as mean ± s.e.m. *p < 0.05 and **p < 0.01