Fig. 4

IHT upregulates the TFEB-related pathway in PAM. A–F Total RNA of in vitro PAM was isolated for RNA-seq. Heatmap (A) and KEGG pathway (B) analyses were used to demonstrate the differentially expressed genes and enriched pathways, respectively. C–F Autophagy-related differential genes, including Lamp1, Vps11, Wipi2, and Vps18, were detected using qRT-PCR. n = 3 (two-way ANOVA). G, H In vitro PAM was stained with an anti-TFEB antibody after a single day of IHT. Scale bar = 20 μm. TFEB signals in the microglial nucleus were quantified. n > 50 (two-way ANOVA). I, J Brain sections of IHT-treated mice were co-stained with anti-Iba1 and anti-TFEB antibodies. Arrows indicate the PAM nucleus. Scale bar = 20 μm. TFEB signals in the PAM nucleus were measured. n > 50 (Student's t test). K–N Autophagy-related genes, such as Lamp1, Vps11, Wipi2, and Vps18, in the brains of IHT-treated mice were detected via qRT-PCR. n = 3 (two-way ANOVA). *P < 0.05, **P < 0.01, and ***P < 0.001. TG, APP/PS1 transgenic mice