Journal: Cell reports

Article Title: Placenta-derived SOD3 deletion impairs maternal behavior via alterations in FGF/FGFR-prolactin signaling axis

doi: 10.1016/j.celrep.2024.114789

Figure Lengend Snippet: (A and G) Time course of prolactin (A) or SOD3 (G) osmotic pump embedding and maternal behavior tests in placental Sod3 KO dam. (B–F and H–L) Plasma levels of prolactin (B and H), number of living pups (C and I), latency of the first retrieval on day 1 (D and J), time spent grooming on day 1 (E and K), and time spent crouching on day 1 (F and L) of WT dams and KO dams with prolactin (B–F) or SOD3 (H–L) osmotic pump embedding of saline or recombinant prolactin. (M–P) mRNA expression levels of prolactin ( Prl ) (M and N) and prolactin receptor ( Prlr ) (O and P) in 200 ng/mL recombinant SOD3-stimulated GH3 cells (M and O) and mouse primary pituitary cells (N and P). N = 5 in each group; three technical replicates for each group; * p < 0.05 and ** p < 0.01.

Article Snippet: Osmotic pumps were filled with 7.5 μg of recombinant mouse prolactin (1445-PL, R&D Systems) or SOD3 diluted in 1 mL of phosphate-buffered saline (PBS).

Techniques: Saline, Recombinant, Expressing

Journal: Cell reports

Article Title: Placenta-derived SOD3 deletion impairs maternal behavior via alterations in FGF/FGFR-prolactin signaling axis

doi: 10.1016/j.celrep.2024.114789

Figure Lengend Snippet: (A) Reactome pathway analysis of placental Sod3 f/f and Sod3 −/− dams. (B) Prl mRNA expression levels of 100 ng/mL recombinant fibroblast growth factor (FGF)1-, FGF2-, or FGF4-stimulated mouse primary pituitary cells with or without 10 μM FGF receptor (FGFR) inhibitors. BGJ398: FGFR1/2/3 inhibitor, PD16686: FGFR1 inhibitor, H3B-6527: FGFR4 inhibitor. N = 5; three technical replicates for each group; ** p < 0.01 vs. FGF1-control, †† p < 0.01 vs. FGF2-control, and ‡‡ p < 0.01 vs. FGF4-control. (C) Prl mRNA expression levels in recombinant FGF1-, FGF2-, or FGF4-stimulated primary pituitary cells from placental Sod3 f/f and Sod3 −/− dams on day 10 after delivery. (D) Phosphorylation levels of FGFR-induced signaling molecules in recombinant FGF1-, FGF2-, or FGF4-stimulated primary pituitary cells from placental Sod3 f/f and Sod3 −/− dams. N = 3 in each group; three biological and technical replicates for each group.

Article Snippet: Osmotic pumps were filled with 7.5 μg of recombinant mouse prolactin (1445-PL, R&D Systems) or SOD3 diluted in 1 mL of phosphate-buffered saline (PBS).

Techniques: Expressing, Recombinant, Control

Journal: Cell reports

Article Title: Placenta-derived SOD3 deletion impairs maternal behavior via alterations in FGF/FGFR-prolactin signaling axis

doi: 10.1016/j.celrep.2024.114789

Figure Lengend Snippet:

Article Snippet: Osmotic pumps were filled with 7.5 μg of recombinant mouse prolactin (1445-PL, R&D Systems) or SOD3 diluted in 1 mL of phosphate-buffered saline (PBS).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Catecholamine ELISA, Sandwich ELISA, Dehydrogenase Assay, Activity Assay, Software

Journal: JCI Insight

Article Title: Radiotherapy exposure directly damages the uterus and causes pregnancy loss

doi: 10.1172/jci.insight.163704

Figure Lengend Snippet: ( A ) Ovariectomized (OVX) adolescent female wild-type mice exposed to 7 Gy TBI (IRR) or nonirradiated controls (Non-IRR) were hormone-primed with estradiol (E2) and a progesterone pellet (P4) before undergoing artificial decidualization. ( B ) Representative images of uteri collected 4 days after artificial decidualization are shown. ( C ) Relative uterine weight (UW) to body weight (BW) was quantified. ( D ) Expression of genes key for decidualization and hormone responses — Esr1 and Pgr — was analyzed by qPCR. ( E ) Cultured primary human endometrial stromal fibroblasts were exposed to 7 Gy γ-irradiation (IRR) or left as nonirradiated controls (Non-IRR) and then artificially decidualized in vitro. ( F ) Representative images highlighting cell morphology between nondecidualized (Non-Decid) and decidualized (Decid) cells are shown. ( G ) The concentration of prolactin in the media was quantified by ELISA. ( H ) Prolactin (PRL) gene expression was assessed by qPCR. Scale bars: 5 mm. Data are mean ± SEM; unpaired t test (2 groups; parametric distribution), Mann-Whitney test (2 groups; nonparametric distribution) or 1-way ANOVA with Holm-Šídák post hoc test; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 2–9/group. Hand2, heart and neural crest derivatives expressed 2.

Article Snippet: Prolactin ELISA was completed according to manufacturer’s instructions (R&D Systems DY682 and DY008).

Techniques: Expressing, Cell Culture, Irradiation, In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: The hyperlipidaemic drug fenofibrate significantly reduces infection by SARS-CoV-2 in cell culture models

doi: 10.1101/2021.01.10.426114

Figure Lengend Snippet: A. Schematic showing ACE2 tagged with LgBIT and SmBIT. B. HEK-293 cells were transfected with combinations of plasmids encoding LgBIT or SmBIT fused to either protein kinase A regulatory subunit (PRKAR2) or catalytic subunit (PRKACA), ATG5 or ACE2. The results (mean ± S.D., n = 5) were normalized to the luminescence measured in cells transfected with protein kinase A reporters (positive control). C. HEK-293 cells were transfected with plasmids encoding ACE2 nanoBIT reporters under the control of the HSV TK promoter and ACE2 or prolactin (PRL) under the control of the CMV promoter. The results (mean ± S.D., n = 4) were normalized to the luminescence measured in cells transfected with protein kinase A reporters and prolactin. D. HEK-293 cells were transfected with NanoBIT-tagged ACE2 reporters and incubated with sodium valproate or clofibrate at a concentration equal to 1x, 2x or 3x the reported C max of the drug. After 1 hour, luminescence was measured and normalized (mean ± S.D., n =4) to that measured in cells treated with DMSO. E. A series of other fibrates were similarly evaluated in the assay. The luminescence measured (mean ± S.D., n = 5-11, solid bars) was significantly different to that measured in cells treated with solvent where shown. When these fibrates were incubated with purified LgBIT and HiBIT-RBD to create a constitutively active nanoluc, each of these fibrates were found to inhibit nanoluciferase (bezafibrate 35 ± 7 %, ciprofibrate 55 ± 6 %, fenofibric acid 46 ± 3 %, fenofibrate 69 ± 5 %, gemfibrozil 61 ± 2 % of the activity measured in the presence of DMSO). To correct for this, the luminescence measurements from cells treated with fibrates in cells were divided by these latter values to estimate the effect of the drugs on dimerization (hatched bars). Significant difference from control is shown as *, P < 0.05; **, P < 0.01; ***, P < 0.005.

Article Snippet: The plasmid pcDNA3 encoding ACE2 was obtained from GenScript (OHu20260); the plasmid encoding prolactin (PRL) was obtained from Sino Biological (HG10275-CY).

Techniques: Transfection, Positive Control, Incubation, Concentration Assay, Purification, Activity Assay