Fig. 3: Golgi PtdIns4P regulates ATGL protein stability in the Golgi apparatus.
From: Glucose controls lipolysis through Golgi PtdIns4P-mediated regulation of ATGL
a, Immunofluorescence of PtdIns4P in HEK293-AAV cells after glucose deprivation (n = 150 cells per group). Scale bar, 5 μm. b, Immunoblot of ATGL in HEK293T cells 72 h after PI4KB and SACM1L knockdown (n = 8 or 4 independent experiments for control or other groups). c, Immunoblot of ATGL in HEK293T cells 24 h after treatment with 5 μM PIK93, 10 μM BF738735 and 10 μM UCB9608 (n = 6 independent experiments per group). d, Immunofluorescence of ATGL in HEK293-AAV cells 24 h after UCB9608 treatment (n = 61 cells per group). Scale bar, 5 μm. e, Immunoblot of ATGL in the different fractions from HEK293T cells 6 h after UCB9608 treatment (n = 6 independent experiments per group). f, Immunoblot of ATGL in HepG2 cells 72 h after PI4KB and SACM1L knockdown (n = 8 or 4 independent experiments for control or other groups). g, Immunoblot of ATGL in HepG2 cells 24 h after treatment with PIK93, BF738735 and UCB9608 (n = 6 independent experiments per group). h,i, Immunoblot of ATGL and determination of basal lipolysis 72 h after knocking down Pi4kb in the iBAs (n = 8 or 4 independent experiments for control or other groups in h; n = 6 independent experiments per group in i). j,k, Immunoblot of ATGL and determination of basal lipolysis 72 h after knocking down Sacm1l in the iBAs (n = 6 or 4 independent experiments for control or other groups in j; n = 6 independent experiments per group in k). l,m, Immunoblot of ATGL (l) and determination of basal lipolysis (m) 24 h after treating iBAs with 1 μM PIK93, 1 μM BF738735 and 1 μM UCB9608 (n = 6 independent experiments per group). Data are presented as mean ± s.e.m. and analysed using two-tailed paired t-test (e), two-tailed Mann–Whitney test (a and d), one-way ANOVA method with Dunnett correction for multiple comparisons between control and other groups (b, c, f–k and m) and Kruskal–Wallis test with Dunn’s correction for multiple comparisons between control and other groups (l). Source numerical data and unprocessed blots are available in .
