Journal: The FASEB Journal

Article Title: Testosterone Exposure During Fetal Masculinization Programming Window Determines the Kidney Size in Adult Mice

doi: 10.1096/fj.202500761RR

Figure Lengend Snippet: Fetal testosterone supplementation rescues the adult kidney size by normalizing gene expression in the fetal kidney. (A) Anogenital distance of 3‐month‐old Hsd17b3 −/− (KO) and WT males without and with testosterone supplementation (res) at E14.5–17.5. The lines indicate means. * = p ≤ 0.05 ** = p ≤ 0.01. (B) kidney weights of 3‐month‐old Hsd17b3 −/− (KO) and WT males without and with testosterone supplementation (res) at E14.5–17.5. The lines indicate means. * = p ≤ 0.05 ** = p ≤ 0.01. (C) Igfbp5 expression (RNA‐seq) in E16.5 kidneys of WT, Hsd17b3 −/− (KO) and treated KO (res) groups (RPKM = Reads Per Kilobase Million). The lines indicate means. * = p ≤ 0.05. (D) Igfbp5 mRNA expression (qPCR) at different age points in WT kidneys. Points indicate means and whiskers SD. d = day, wk. = week. (E) Representative western blot images in E18.5 and quantification of IGFBP5 and loading control β‐actin in E15.5 and E18.5 WT and Hsd17b3 −/− (KO) kidney homogenates. The lines indicate means. * = p ≤ 0.05. (F) Immunofluorescent staining of IGFBP5 (red) and LRP2 (green) in E18.5 and 2‐week‐old WT and Hsd17b3 −/− (KO) kidneys. Scale bar: 50 μm.

Article Snippet: Cells were cultured on poly‐L‐lysine–coated plates (#P4832, Sigma‐Aldrich) and switched to serum‐free medium (DMEM/F12, #D2906, Sigma; 1% Penicillin/streptomycin, #A5256701, Gibco; 1% L‐Glutamine, #25030–024, Gibco; 20 nM DHT) for 3 h before changing to fresh serum‐free medium with 25, 50, or 100 nM recombinant IGFBP5 (#578‐B5, R&D Systems).

Techniques: Gene Expression, Expressing, RNA Sequencing, Western Blot, Control, Staining

Journal: The FASEB Journal

Article Title: Testosterone Exposure During Fetal Masculinization Programming Window Determines the Kidney Size in Adult Mice

doi: 10.1096/fj.202500761RR

Figure Lengend Snippet: IGFBP5 is likely regulated by AR and HNF4A, and in turn can affect cell proliferation. (A) Visualization of AR and HNF4A binding peaks and H3K4m1 and H3K27ac histone marks near Igfbp5 in ChIP‐seq data from Pihlajamaa et al., 2014. T = testosterone treatment. (B) Proliferation analysis of HEK 239 cells cultured with 0 (Ctrl), 25, 50, or 100 nM recombinant human IGFBP5. The lines indicate means. * = p ≤ 0.05. (C) Representative western blot images in E15.5 and quantification of phosphorylated AKT (pAKT), AKT, and loading control β‐actin in E15.5 and E18.5 WT and Hsd17b3 −/− (KO) kidney homogenates. The lines indicate means. (D) Representative western blot images in E15.5 and quantification of phosphorylated S6 ribosomal protein (pS6) and loading control β‐actin in E15.5 and E18.5 WT and Hsd17b3 −/− (KO) kidney homogenates. The lines indicate means. ** = p ≤ 0.01.

Article Snippet: Cells were cultured on poly‐L‐lysine–coated plates (#P4832, Sigma‐Aldrich) and switched to serum‐free medium (DMEM/F12, #D2906, Sigma; 1% Penicillin/streptomycin, #A5256701, Gibco; 1% L‐Glutamine, #25030–024, Gibco; 20 nM DHT) for 3 h before changing to fresh serum‐free medium with 25, 50, or 100 nM recombinant IGFBP5 (#578‐B5, R&D Systems).

Techniques: Binding Assay, ChIP-sequencing, Cell Culture, Recombinant, Western Blot, Control

Journal: Journal of the Endocrine Society

Article Title: Subclinically low BMD in young men is associated with compromised bone microarchitecture and lower lean mass

doi: 10.1210/jendso/bvag003

Figure Lengend Snippet: Biomarkers. Fig. 1 depicts all assessed biomarker concentrations. The total hip, femoral neck, or lumbar spine (Lower) group is depicted in black filled circles, while the control (Con) group is in clear squares. A, P1NP (pg/mL); B, alkaline phosphatase (ALP) (pg/mL); C, tartrate-resistant acid phosphatase 5b (TRAP5b) (U/L); D, sclerostin (pmol/L); E, β C-terminal cross-linked telopeptide of type I collagen (βCTX) (pmol/L); F, testosterone (ng/mL); G, estradiol (pg/mL); H, parathyroid hormone (PTH) (pg/mL); I, insulin-like growth factor-1 (IGF-1) (ng/mL); J, insulin-like growth factor-1 binding protein 5 (IGFBP-5) (ng/mL).

Article Snippet: Commercially available enzyme-linked immunosorbent assays were used to determine circulating concentrations of serum procollagen type I N-terminal propeptide (PINP; Novus catalog No. NBP2-76465, RRID:AB_3720306, 1:200 dilution, CV 5.3%, sensitivity 9.4 pg/mL), alkaline phosphatase (ALP; Novus catalog No. NBP2-68197, RRID:AB_3720307, CV 6.0%, sensitivity 46.9 pg/mL), tartrate-resistant acid phosphatase 5b (TRAP5b; Immunodiagnostic Systems catalog No. SB-TR201A, RRID:AB_3095995, CV 4.7%, sensitivity 0.4 U/L), sclerostin (Biomedica catalog No. BI-20492, RRID:AB_2894889, CV 6.4%, sensitivity 3.2 pmol/L), testosterone (Alpco Diagnostics catalog No. 11-TESHU-E01, RRID:AB_3720308, CV 4.2%, sensitivity 0.02 ng/mL), estradiol (Alpco Diagnostics catalog No. 11-ESTHU-E01, RRID:AB_2756385, CV 7.6%, sensitivity 10 pg/mL), insulin like growth factor-1 (IGF-1; Alpco Diagnostics catalog No. 22-IGFHU-E01, RRID:AB_2923276, 1:20 dilution, average CV 3.3%, assay sensitivity 9.4 pg/mL), IGF binding protein-5 (IGFBP-5; R and D Systems catalog No. DY875, RRID:AB_3720311, 3× dilution, CV 2.7%), parathyroid hormone (PTH; (Alpco Diagnostics catalog No. 21-IPTHU-E01, RRID:AB_2943035), CV 7.3%, sensitivity 1.6 pg/mL), and plasma β C-terminal cross-linked telopeptide of type I collagen (βCTX; Immunodiagnostic Systems catalog No. IS-3000N, RRID:AB_3720313, CV 6.2%, sensitivity 3.2 pmol/L).

Techniques: Biomarker Discovery, Control, Binding Assay