This document provides an overview of oxygen (O2) levels and their influence on cell culture experiments. It defines terms like hypoxia, normoxia, and hyperoxia in relation to different tissue oxygen levels. The document discusses how O2 levels inside a standard CO2 incubator create hyperoxic conditions for cells, as the O2 partial pressure is 141 mmHg compared to physiological levels of 40-100 mmHg in tissues. It also summarizes the hypoxia-inducible factor 1 (HIF-1) signaling pathway, which senses oxygen levels in cells and regulates gene expression. The takeaway messages are that researchers should carefully define and control O2 levels in experiments to match physiological conditions and improve reproducibility.