**3.1 Methods**

NGF synthesis was measured as described by Hazekawa et al. [37]. Male ddY mice (25–30 g weight) obtained from Kiwa Laboratory Animals Co., Ltd. (Wakayama, Japan), were housed under a 12-h light/dark cycle at room temperature and 55 ± 5% humidity. The lyophilized mycelia from *H. erinaceum* and *H. ramosum* were suspended in purified water and the samples were orally administrated to mice once a day for 14 days. NGF levels were analyzed in the cortex, striatum, and hippocampus 24 h after the last administration. Results are expressed as mean ± standard error of mean (SEM). The standard dose (300 mg/kg body weight) was determined based on Hazekawa et al. [37]. More detailed methodology can be found in Suruga et al. [9].

## **3.2 Stimulation of NGF synthesis by** *H. ramosum* **and** *H. erinaceum* **mycelia**

The effects of 14-days of oral administration of 300 mg/kg *H. erinaceum* mycelia and *H. ramosum* mycelia on NGF levels in intact mouse brains are shown in **Figure 4**. *H. ramosum* mycelia were more potent than *H. erinaceum* mycelia in terms of NGF stimulation in the hippocampus of intact mice. Processing of *H. ramosum* mycelia over time significantly increased NGF levels in the hippocampus.

Different regions of the mouse brain responded differently to application of varying concentrations of *H. ramosum* mycelia on NGF synthesis [9]. The NGF levels in hippocampus increased with increased concentration of *H. ramosum* mycelia, while such dose-dependent response was not seen in cortex or striatum (**Figure 5**).
