Fig. 2

JQ1 positively regulates FBP1 protein stability in PDAC. a-d, PANC-1 and SW1990 cells were treated with different dose of JQ1 as indicated. After 24 h, cells were harvested for western blot analysis (a), RT-qPCR (b). Data are shown as means ± SD (n = 3). n.s., not significant. Measurement of glucose consumption (c) and L-lactate production (d) in the spent medium of PANC-1 cells treated with different dose of JQ1 as indicated, medium was collected after 24 h. Data are shown as means ± SD (n = 3). *, P < 0.05. e-h, PANC-1 and SW1990 cells were treated with 2 μM of JQ1. After different time points as indicated, cells were harvested for western blot analysis (e) and RT-qPCR (f). Data are shown as means ± SD (n = 3). n.s., not significant. Measurement of glucose consumption (g) and L-lactate production (h) in the spent medium of PANC-1 cells treated with 2 μM of JQ1, medium was collected in different time point as indicated after JQ1 treatment. Data are shown as means ± SD (n = 3). *, P < 0.05. i and j, PANC-1 cells were treated with different dose of JQ1 as indicated. After 24 h, cells were treated with 50 μg/μl cycloheximide (CHX). At different time points, cells were harvested for western blot analysis. At each time point, the intensity of FBP1 was normalized to the intensity of β-Tubulin (loading control) first and then to the value at the 0-h time point. k, PANC-1 cells were transfected with the indicated constructs. 24 h post transfection, cells were treated with different dose of JQ1 as indicated for another 24 h,then cells were harvested for western blot analysis. Cells were treated with or without 20 μM of MG132 for 8 h before harvested. l, PANC-1 cells were transfected with the indicated constructs. 24 h post-transfection, cells were treated with 2 μM of JQ1. After different time points as indicated, cells were harvested for western blot analysis. Cells were treated with or without 20 μM of MG132 for 8 h before harvested