Journal: The Journal of Biological Chemistry

Article Title: Peptide OM-LV20 protects astrocytes against oxidative stress via the ‘PAC1R/JNK/TPH1’ axis

doi: 10.1016/j.jbc.2022.102429

Figure Lengend Snippet: OM-LV20 inhibited phosphorylation of MAPK signaling pathway and increased levels of TPH1 and PAC1R in CTX-TNA2 cells under H 2 O 2 stimulation. A , representative Western blots of MAPK, TPH1, and PAC1R protein levels in CTX-TNA2 cells. B , OM-LV20 significantly promoted mRNA level of PAC1R. C – F , histogram of PAC1R, TPH1, p-JNK, and p-p38 protein expression, respectively. Data are means ± SEM. n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Primary antibodies included the following: GAPDH (1:3000; Proteintech); p-JNK, JNK, p-p38, and p38 (1:2000; Cell Signaling); TPH1 (1:500; Abcam); and PAC1R (1:500; ABclonal).

Techniques: Phospho-proteomics, Western Blot, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Peptide OM-LV20 protects astrocytes against oxidative stress via the ‘PAC1R/JNK/TPH1’ axis

doi: 10.1016/j.jbc.2022.102429

Figure Lengend Snippet: OM-LV20 regulated TPH1 by binding to PAC1R in CTX-TNA2 cells. A , yellow represents PAC1R conformation, and blue represents OM-LV20 conformation. PAC1R and OM-LV20 showed good shape complementarity. B , yellow is residue of OM-LV20, and blue is PAC1R protein residue. LEU1 formed hydrogen bonds with receptor proteins SER23 and ASN60; LYS4 formed hydrogen bonds with receptor protein ILE19; ALA9 formed hydrogen bonds with TYR118. C , yellow is hydrophobic residue of OM-LV20, and blue is hydrophobic residue of PAC1R protein. They showed hydrophobic accumulation and Van der Waals interactions. D , application of PACAP6-38 partly weakened increase in TPH1 by OM-LV20. E , histogram of TPH1 protein expression. Data are means ± SEM. n = 7, ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Primary antibodies included the following: GAPDH (1:3000; Proteintech); p-JNK, JNK, p-p38, and p38 (1:2000; Cell Signaling); TPH1 (1:500; Abcam); and PAC1R (1:500; ABclonal).

Techniques: Binding Assay, Residue, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Peptide OM-LV20 protects astrocytes against oxidative stress via the ‘PAC1R/JNK/TPH1’ axis

doi: 10.1016/j.jbc.2022.102429

Figure Lengend Snippet: Neuroprotective mechanism of OM-LV20 in astrocytes. Peptide OM-LV20 exerts protective effects on astrocytes via the ‘PAC1R/JNK/TPH1’ axis to resist oxidative stress.

Article Snippet: Primary antibodies included the following: GAPDH (1:3000; Proteintech); p-JNK, JNK, p-p38, and p38 (1:2000; Cell Signaling); TPH1 (1:500; Abcam); and PAC1R (1:500; ABclonal).

Techniques:

Journal: Translational Psychiatry

Article Title: The ventral hippocampus to paraventricular thalamus circuit regulates context-dependent hyperlocomotion through PAC1 receptor signaling in the chronic stress-induced PTSD mouse model

doi: 10.1038/s41398-026-03963-1

Figure Lengend Snippet: (A) The immunofluorescent staining showed that CSDS treatment induced obvious increases in the expression of PAC1 receptor within PVT neurons (upper panel, amplification: 4 x; lower panel, amplification: 60 x). (B) The western blot revealed the apparent increases in the PAC1 receptor within PVT region and (C) the statistical analysis ( n = 5 mice for each group; Mann-Whitney unpaired two-tailed U test; U = 0, ** p = 0.0079). (D) The experimental strategy of cannula injection of PAC1 receptor antagonist and behavioral tests. (E) The context-dependent locomotion activity (One-way ANOVA with Tukey post-hoc test, F(2, 21.47) = 9.201, ** p = 0.0013; Ctrl vs. CSDS, * p = 0.0429; CSDS vs. CSDS + PA-915, *** p = 0.0006). (F) The time spent in open arm (One-way ANOVA with Tukey post-hoc test, F(2, 24.69) = 9.033, ** p = 0.0011; Ctrl vs. CSDS, ** p = 0.0041; CSDS vs. CSDS + PA-915, * p = 0.0163) and (G) number of open arm entries (One-way ANOVA with Tukey post-hoc test, F(2, 26.45) = 6.594, ** p = 0.0047; Ctrl vs. CSDS, * p = 0.0107; CSDS vs. CSDS + PA-915, * p = 0.0193) in the elevated plus maze test. (H) The sociability in the three-chamber test (One-way ANOVA with Tukey post-hoc test, F(2, 17.81) = 6.317, ** p = 0.0084; Ctrl vs. CSDS, * p = 0.0189; CSDS vs. CSDS + PA-915, * p = 0.0173) and (I) the innate motivation in the female encounter test (One-way ANOVA with Tukey post-hoc test, F(2, 22.87) = 7.469, ** p = 0.0032; Ctrl vs. CSDS, * p = 0.0122; CSDS vs. CSDS + PA-915, * p = 0.0102). (J) The arousal levels (One-way ANOVA with Tukey post-hoc test, F(2, 24.94) = 5.412, * p = 0.0112; Ctrl vs. CSDS, * p = 0.044; CSDS vs. CSDS + PA-915, * p = 0.0316). Data was represented as mean ± SD from three independent experiments. n = 10 mice per group per experiment.

Article Snippet: The diluted primary antibodies against PAC1R (CST, 1:1000) and GADPH (CST, 1:3000) were introduced, and the mixture was incubated for 12 h at 4 °C.

Techniques: Staining, Expressing, Amplification, Western Blot, MANN-WHITNEY, Two Tailed Test, Injection, Activity Assay