Proliferation and AMPK phosphorylation of shControl/shATGL cancer cells. (A) Protein expression levels of ATGL, HSL, and MGL in shControl/shATGL cancer cell lines were analyzed by western blot analysis. GAPDH was used as housekeeping protein. Equal protein loading was also verified by Coomassie blue staining of the membrane. (B-D) Proliferation of shControl/shATGL (B) C26, (C) HepG2, and (D) LLC cells was analyzed using a hemocytometer at the indicated timepoints. (E) P-AMPK (Thr172) and AMPK protein abundance in shControl/shATGL cancer cells was analyzed by western blot analysis. GAPDH was used as loading control. (F) Relative phosphorylation level of AMPK in shControl/shATGL cancer cells was determined by densitometric quantification of the immunoblots. (G) ATGL mRNA expression in murine cancer cell lines and MEFs was analyzed by qPCR. (H) ATGL protein abundance in murine cancer cell lines and MEFs was analyzed by western blot analysis. GAPDH was used as loading control. (I) Relative protein expression level of ATGL in mouse cancer cells and MEFs was determined by densitometric quantification of the immunoblots. Data are presented as mean values of n = 3–6 ± standard deviation. Significance was determined by student's t-test (*p ≤ 0.05, **p ≤ 0.01).