Fig 2: Increased phosphorylated-cPLA2 in APOE4 is mediated by p38 MAPK. A Phosphorylated and total p38 and ERK levels in primary astrocyte from ApoE3 and ApoE4-TR mice were detected by WB (n = 3 for each genotype). B Phosphorylated and total p38 and ERK levels in cortical homogenates from ApoE3 and ApoE4-TR mice were detected by WB. (n = 5 for each genotype). C ApoE4 primary astrocytes from mouse were pre-treated with p38 inhibitors SB 202190 (10 μM) or SB 203580 (10 μM) for 20 minutes and then treated with medium or TNFα plus IFNγ together for 30 minutes. The total and phosphorylated cPLA2 and p38 were detected in the cell lysate by WB. Quantification was obtained from 3 independent repeats. D cPLA2 bound with p38. Immunoprecipitation was performed in the cell lysate of immortalized ApoE4 astrocytes using anti-cPLA2 antibody or species-matched IgG. cPLA2 and p38 were co-detected after immunoprecipitation by WB. WB: Western Blot. Data are represented as mean ± SEM and analyzed by Student’s t-test (two-tailed). *p < 0.05

Fig 3: Study workflow and clinical-pathological and molecular characteristics of SYN+ NSCLCs(A) Diagram capturing the steps of the immuno-LCM workflow developed for this study. SYN+ clones were first identified using standard IHC. SYN+ and SYN− clones were isolated under direct visualization using an infrared (IR)-based LCM system. Expression/activation of 57 signal transduction molecules were measured on a continuous scale using the RPPA and compared across different clones to identify signaling events that were uniquely attributable to SYN+ cells.(B) Examples of SYN+ cells identified by IHC in a primary tumor (left site) and a lymph node metastasis (right panel) in patients with NSCLC; SYN+ cells differ in degrees of distribution: from a few to 70% of metastatic cells.(C) Higher rates of distant metastases (p = 0.03), recurrence (p = 0.03), and poorer prognosis (p = 0.05) were detected in patients whose tumor contained SYN+ cells. Bar graph illustrating frequency of distant metastases and outcome of patients with NSCLC with and without SYN+ neuroendocrine-like cells.(D) Bar graphs displaying changes in SYN expression in microdissected matched SYN+ and SYN− cancer cells. IHC intensity staining for each sample is reported along with the RPPA-based fold change values.(E) Selected examples of IHC staining matching the samples displayed in the bar graph in (D).(F) Boxplot illustrating significant changes in SYN expression between matched SYN+ and SYN− microdissected clones (p = 0.02).

Fig 4: Intratumor heterogeneity analyses reveal different extents of NE differentiation.A UMAP visualization of epithelial cell sub-clusters from each sample. B Heatmap depicting prostate lineage marker genes and AR pathway gene expression levels in epithelial cell sub-clusters from each sample. Those highlighted in red frame showed cluster 5 in patient #4 and cluster 4 in patient #6 was NE sub-clusters. C Immunohistochemistry (IHC) staining for K5, K18, AR, SYP, and SOX2 in sections from five samples. Scale bars represent 50 μm.

Fig 5: Effect of Dio on synaptic loss in young AppNL-G-F mice. (A–C) Representative Golgi-staining and quantification showing spines of 3- and 6-month-old male and female WT and AppNL-G-F mice. Scale bar: 0.1 mm. Data are expressed as mean ± SEM (n = 8). * p < 0.05 compared with WT; ** p < 0.01 compared with WT. # p < 0.05 compared with AppNL-G-F; (F,K) TEM image from male and female WT and AppNL-G-F mice showing synapses. Scale bar: 0.5 μm; (F) 3-month-old male and female mice; (K) 6-month-old male and female mice. (D,E,I,J) Immunofluorescence staining of PSD95 and SYP in the cortex and hippocampus in 3- and 6-month-old male and female WT and AppNL-G-F mice. Scale bar: 0.1 mm; (D) 3-month-old male mice; (E) 3-month-old female mice; (I) 6-month-old male mice; (J) 6-month-old female mice. (G,H,L,M) Western blot and quantification of PSD95 and SYP protein expression in the cortex and hippocampus in 3- and 6-month-old male and female WT and AppNL-G-F mice. Data are expressed as mean ± SEM. n = 6 mice/group. * p < 0.05 compared with WT; ** p < 0.01 compared with WT; *** p < 0.001 compared with WT. # p < 0.05 compared with AppNL-G-F; ## p < 0.01 compared with AppNL-G-F.