Fig. 5
From: Caspase-1 cleaves PPARγ for potentiating the pro-tumor action of TAMs
Loss of function of PPARγ fails to promote TAM differentiation or metabolic reprogramming in TAMs. a–d THP-1 cells stably expressing control scramble shRNA were cultured alone (Ctrl + scr) or cocultured with MCF-7 (Coculture + scr), THP-1 cells stably expressing shRNA against PPARγ were cultured alone (Ctrl + shPPARγ) or cocultured with MCF-7 (Coculture + shPPARγ), after 2 d, TAM hallmarks expression was analyzed by RT-qPCR (a), THP-1 macrophages were stained with Oil Red O and lipid content was quantified (b), or MCF-7 and regular culture medium was replaced with 2 ml fresh culture medium then culture for another 24 h, the lactate concentration were measured (c), PPARγ and its fragment were analyzed by immunoblot (d). e-h THP-1 cells stably expressing LacZ were cultured alone or cocultured with MCF-7, PPARγ knockout THP-1 cells or THP-1 cells that express PPARγ (D64A) were cocultured with MCF-7, after 2 d, TAM hallmarks expression was analyzed by RT-qPCR (e), THP-1 macrophages were stained with Oil Red O and lipid content was quantified (f), or MCF-7 and regular culture medium was replaced with 2 ml fresh culture medium then culture for another 24 h, the lactate concentration was measured (g), PPARγ and its fragment were analyzed by immunoblot (h). i, j THP-1 macrophages were cultured alone (Ctrl) or cocultured with MCF-7 (Coculture) in the presence or absence of PPARγ inhibitor GW9662. After 2 days, THP-1 macrophages were stained with Oil Red O and lipid content was quantified (i), or MCF-7 and regular culture medium was replaced with 2 ml fresh culture medium then culture for another 24 h, the lactate concentration was measured (j). All the histograms in this figure show means ± s.e.m. (n = 3, *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant, one way ANOVA for multiple comparisons)
