Journal: The Journal of Clinical Investigation

Article Title: Chloride homeostasis dysfunction drives hyperactivation of corticotropin-releasing factor-expressing neurons in the amygdala in stress-induced hypertension

doi: 10.1172/JCI195536

Figure Lengend Snippet: ( A and B ) Changes in MAP ( A ) and HR ( B ) measured by radiotelemetry before, during and after CUMS in WKY rats, BHRs and age-matched unstressed WKY rats and BHRs ( n = 6 rats in each group). Data are expressed as means ± SEM. * P < 0.05 for CUMS BHR versus unstressed BHRs, # P < 0.05 for CUMS BHR versus CUMS WKY, ^ P < 0.05 for CUMS WKY versus unstressed WKY. Repeated measures of 2-way ANOVA with Tukey’s post hoc test. ( C ) Representative immunofluorescence images showing CRF immunopositive neurons (green), delta-FosB immunopositive neurons (red), and CRF and delta-FosB double-positive neurons in the CeA of WKY, CUMS WKY, BHR, and CUMS BHR group of rats ( n = 5 rats in each group). ( D and E ) Summary data showing the numbers of CRF-positive neurons ( D ) and delta-FosB positive neurons ( E ) in the CeA of these 4 groups. ( F ) The percentage of CRF and delta-FosB double-positive neurons in the CeA of these 4 groups ( n = 5 rats in each group). Data are expressed as means ± SEM. ** P < 0.01, *** P < 0.001. One-way ANOVA with Tukey’s multiple comparison tests.

Article Snippet: Subsequently, the sections were incubated with a mouse anti-delta-FosB antibody (1:50, #sc-398595), a rabbit anti-CRF antibody (1:100, #A1122, Abclonal) or a rabbit anti-NKCC1 antibody (1:100, #13884, Proteintech) overnight at 4°C.

Techniques: Immunofluorescence, Comparison

Journal: BMC Medicine

Article Title: Identification of cryosensitive niches and a targetable FOS/AP‑1 program in the human ovarian cortex by single‑cell and spatial transcriptomics

doi: 10.1186/s12916-026-04757-4

Figure Lengend Snippet: FOS/AP-1 is rapidly activated in the early flow of vitrification, and its inhibition maintains the viability of frozen-thawed ovaries in vitro. A The circle plots demonstrated the cross talk between PCs (left)/GCs (right) and other cell types in fresh and vitrification-thawed subjects respectively. B Dot plot of selected ligand/receptor (left) and receptor/ligand pair(right) interactions between FOS + PCs or FOS/AP-1 active GCs and other cell components, analyzed separately for fresh and vitrification-rapid warming groups. C Regulon specificity scores (RSSs) identified top specific regulons in fresh and vitrification-rapid warming GCs. The top 6 regulons from each group are marked in red and annotated. D Circle plot shows gene regulatory network of the TFs FOS and JUND and their target genes. E Immunofluorescence staining was employed to detect the expression changes of EGR1 in human ovary tissue following vitrification-rapid warming treatment. Scale bar = 200 μm. F Schematic diagram illustrating the freezing and thawing process of mouse ovarian tissue, with corresponding protein collection points for G . G Western blotting of AP-1 complex family (FOS, FOSB, and JUN) and the upper stream protein (p-AKT and AKT) and down streams (EGR1) expression changes during the whole ovarian vitrification and thawing procedure. The protein collection time point is as follows: A Fresh ovary (FO), B vitrification dehydration (VD), C liquid nitrogen freezing (F), D thawing (T). H Whole-mount immunofluorescence was used to assess vascular (CD31) and extracellular matrix (αSMA) maintenance in in vitro cultured mouse ovarian tissues across different treatment groups. I TUNEL assay of ovaries from fresh and frozen-thawed and T-5224-treated group. Data are presented as the mean ± SEM. n = 5 for each group (unpaired two-tailed t -test). J Schematic illustration showing that activation of FOS/AP-1 in PCs and GCs cells contributes to ovarian cryoinjury-induced apoptosis

Article Snippet: The primary antibodies utilized for these experiments were as follows: PTGDS (1:200, ab182141, Abcam), EGR1 (10 μg/ml; 55117-1-AP, Proteintech), FOSB (2 μg/ml, 2251 Cell Signal Technology), SOHLH2 (1:100, bs-12279R Biossusa), αSMA (1:500; A5228 Sigma), and DDX4 (2 μg/ml; ab180642 Abcam).

Techniques: Inhibition, In Vitro, Immunofluorescence, Staining, Expressing, Western Blot, Cell Culture, TUNEL Assay, Two Tailed Test, Activation Assay