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: Clinical Proteomics, 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, Phospho-proteomics

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, Clinical Proteomics, Sandwich ELISA, Dehydrogenase Assay, Activity Assay, Software

Journal: Cancer research

Article Title: Asporin Restricts Mesenchymal Stromal Cell Differentiation, Alters the Tumor Microenvironment, and Drives Metastatic Progression

doi: 10.1158/0008-5472.CAN-18-2931

Figure Lengend Snippet: ASPN binds to BMP-4 and restricts BMP-4-induced signaling. A, GSEA of TGF-β family pathway genes in Aspn+/+ and Aspn−/− fetal MSCs as determined from microarray (n=3). B, C, BMP-4-induced signaling in Aspn+/+ and Aspn−/− fetal MSCs (A and B represent independently derived MSCs) as measured by immunoblotting and quantified using ImageJ. Aspn+/+ and Aspn−/− fetal MSCs were serum-depleted and then incubated with recombinant mouse BMP-4 (5ng/mL) for 30 minutes. Statistical analyses performed using Student’s t-test (n≥3). D, E, Recombinant mouse ASPN restricts BMP-4-induced signaling in Aspn−/− fetal MSCs as measured by immunoblotting and quantified using ImageJ. Aspn+/+ and Aspn−/− fetal MSCs were serum-depleted and then incubated with recombinant mouse BMP-4 (5ng/mL) with vehicle or recombinant mouse ASPN (100ng/mL) for 30 minutes. Statistical analyses performed using Student’s t-test (n≥2). F, G, BMP-4-induced signaling in WPMY-1-Neo, WPMY-1-ASPN D13, and WPMY-1ASPN D14 (A and B represent independently derived clones) as measured by immunoblotting and quantified using ImageJ. WPMY-1-Neo, WPMY-1-ASPN D13, and WPMY-1ASPN D14 were serum-depleted and then incubated with recombinant human BMP-4 (5ng/mL) for 30 minutes. Statistical analyses performed using one-way ANOVA with Tukey multiple comparison (n≥3). H, Co-immunoprecipitation of ASPN and BMP-4. FLAG tagged mouse ASPN was transfected in HEK293T cells and then incubated with recombinant mouse BMP-4. ASPN was immunoprecipitated with anti-FLAG beads and then examined by immunoblotting for ASPN and BMP-4 (n=2). I, Aspn+/+ and Aspn−/− fetal MSCs were cultured in osteogenic-inducing media with either vehicle, 250nM LDN-193189, or recombinant mouse BMP4 (5ng/mL) and then stained for Alizarin Red. J, Aspn+/+ and Aspn−/− fetal MSCs were cultured in osteogenic-inducing media with either vehicle, 250nM LDN-193189, or recombinant mouse BMP4 (5ng/mL) and then examined for osteogenic-induced genes by qRT-PCR. Statistical analyses performed using one-way ANOVA with Tukey multiple comparison (n≥3). Graphs shown as mean±SEM, *P≤0.05, **P≤0.01, ***P≤0.001, and black bars=100μM.

Article Snippet: Co-immunoprecipitation assays HEK293T cells were transfected with a murine ASPN-Myc-FLAG tagged expression vector (Origene) or vehicle control using Lipofectamine 3000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

Techniques: Microarray, Derivative Assay, Western Blot, Incubation, Recombinant, Clone Assay, Immunoprecipitation, Transfection, Cell Culture, Staining, Quantitative RT-PCR

Journal: Cancer research

Article Title: Asporin Restricts Mesenchymal Stromal Cell Differentiation, Alters the Tumor Microenvironment, and Drives Metastatic Progression

doi: 10.1158/0008-5472.CAN-18-2931

Figure Lengend Snippet: Key Resources

Article Snippet: Co-immunoprecipitation assays HEK293T cells were transfected with a murine ASPN-Myc-FLAG tagged expression vector (Origene) or vehicle control using Lipofectamine 3000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

Techniques: Plasmid Preparation, Magnetic Beads, Recombinant, Transfection, Staining, Generated, Blocking Assay, Bicinchoninic Acid Protein Assay, Cell Migration Assay, Mutagenesis, Random Hexamer Labeling, Expressing, Software