Figure 1. Crystalline silica and low dose LPS enhance glycolysis without affecting macrophage viability.

RAW 264.7 macrophages were exposed to LPS (1 ng/ml) or silica (50 μg/cm²) or primed with LPS (1 ng/ml for 24h) and then exposed to silica (50 μg/cm²) and studied at time points of T0, 2h, 4h, 6h, 24h. The cell supernatants were examined for the concentration of released (A–B) lactate and (C) LDH. (D–E) Annexin V and Propidium iodide were used to analyze cell survival by flow cytometry: (D) graph summarizing the survival of cells treated with LPS (10 ng/ml) or silica (50 μg/cm²) or primed with LPS (1 ng/ml for 24h) and then exposed to silica (50 μg/cm²). (E) Gating strategy to differentiate cell death by early apoptosis (Annexin V⁺PI⁻), late apoptosis (Annexin V⁺PI⁺), or necrosis (Annexin V⁻/PI). Data were normalized by equalizing the protein concentration or equating the actual number of live cells used to perform the experiments. N=3 **** p<0.0001