Fig. 6

A Western blot analysis for HIF-1 α, HSP70, Cathepsin B and Cathepsin L with β-actin in protein lysate collected from CAl33 spheroids with and without macrophages on day 3, 5, 7 and 9, B-E Densitometry analysis for HIF-1 α, HSP70, Cathepsin B and Cathepsin L in western blots normalized with β-actin as a positive control (n = 2), F Western blot analysis for HIF-1 α, HSP70, Cathepsin B and Cathepsin L with β-actin in protein lysate collected from 25,000 M0 macrophages treated with CoCl2 at 0 μM, 50 μM, 100 μM, and 150 μM for 48 h to induce hypoxia to mimic condition like spheroid with macrophage. G-J Real time PCR analysis for HIF-1 α, HSP70, Cathepsin B and Cathepsin L in mRNA isolated from CAl33 spheroids with and without macrophages on day 3, 5, 7 and 9. The western blots were performed for all the conditions in duplicate (n = 2, with 10 spheroids in each condition). Statistical significance was analyzed using two-way ANOVA with Sidak's multiple comparisons test and the p-value obtained are indicated (ns p = 0.5, * p ≤ 0.01, ** p ≤ 0.001, *** p ≤ 0.0001, **** p ≤ 0.00001