β amyloid aβ 1 42  (MedChemExpress)

Journal: Advanced Science

Article Title: Role of Testosterone Signaling in Microglia: A Potential Role for Sex‐Related Differences in Alzheimer's Disease

doi: 10.1002/advs.202413375

Figure Lengend Snippet: Sex‐based differences in microglial autophagy. A) Confocal images of IBA1 (green), p62 (a selective autophagy adaptor protein, magenta), and amyloid β (Aβ, red) in the cerebral cortex of 7‐month‐old 5xFAD mice. Scale bar: 50 µm. The panels on the far‐right show enlarged images of a white square area on the corresponding merged images. Thirty immunohistochemistry image sets were obtained from each of three male or three female mouse cerebral cortices. One representative image set is shown. Three additional sets are shown in Figure (Supporting Information). B,C) Quantification of the number of microglia (B) and Aβ‐positive area (C). Each point represents the number (B) or area (C) per field. D) Pearson's coefficient indicating colocalization of p62 and IBA1. Data are presented as the mean ± SD (B–D, n = 30), ns: not significant. * p < 0.05, *** p < 0.001 by unpaired two‐tailed t‐ test.

Article Snippet: Amyloid β‐protein 1–42 (Aβ 1–42 ) (Peptide Institute Inc., Osaka, Japan) was dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 0.5 m m and stored at ‐20 °C until use.

Techniques: Immunohistochemistry, Two Tailed Test

Journal: Advanced Science

Article Title: Role of Testosterone Signaling in Microglia: A Potential Role for Sex‐Related Differences in Alzheimer's Disease

doi: 10.1002/advs.202413375

Figure Lengend Snippet: Testosterone promotes autophagy by inhibiting the GPRC6A‐mediated ERK/mTOR signaling pathway. A) Relative mRNA expression of Ar (upper) and Gprc6a (lower). Skeletal muscle was used as positive control. B) Western blot analysis of total and phosphorylated forms of ERK in MG6 cells stimulated with testosterone for 1 h at different concentrations. Representative blots (left images) and the relative phosphorylated/total protein ratio for ERK (right graph) are shown. C) Western blot analysis of total and phosphorylated forms of ERK and S6K1 in MG6 cells stimulated with 20 n m of testosterone for the indicated times. Representative blots (left images) and the relative phosphorylated/total protein ratio for ERK and S6K1 (right graphs) are shown. D) GPRC6A was silenced in MG6 cells using shRNAs #1 and #2 and treated with 100 n m testosterone for 2 h. Representative blots and the relative phosphorylated/total for ERK are shown. E) FACS analysis of control and GPRC6A‐silenced MG6 cells treated with HiLyte Fluor 488‐labeled Aβ 1‐42 for 12 h, with or without testosterone added 1 h before Aβ 1–42 stimulation. Analyses were repeated five times. Data are presented as the mean ± SD (A and D, n = 3; B, C, and E, n = 5). ns: not significant. * p < 0.05, for indicated comparisons, by one‐way ANOVA followed by Tukey‐Kramer's HSD test (B,C) or by unpaired two‐tailed t‐ test (D,E).

Article Snippet: Amyloid β‐protein 1–42 (Aβ 1–42 ) (Peptide Institute Inc., Osaka, Japan) was dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 0.5 m m and stored at ‐20 °C until use.

Techniques: Expressing, Positive Control, Western Blot, Control, Labeling, Two Tailed Test