John, N. S.; Tang, K. S.; Urman, M. A.; Lee, C.

Temperature is a critical factor that modulates cellular metabolism and stem cell regulation. Despite extensive studies, the influence of temperature on stem cell regulation via Notch signaling has been limited to studies relying on studies that involve indirect readouts to Notch activation. This study systematically analyzes the effects of temperature on the Notch signaling transcriptional response at the chromosomal, cellular, and tissue levels. Using complementary direct Notch readouts, we demonstrate that Notch activation remains largely unchanged across temperatures, suggesting the presence of temperature-compensatory mechanisms that maintain robust Notch activation. Notch transcriptional activity readouts, however, increased with temperature, indicating that elevated temperatures may enhance Notch transcriptional activity at the chromosomal level. These findings provide a comprehensive framework for understanding effects of temperature and offer new insights into the regulation of Notch signaling in stem cell biology.