Journal: Clinics

Article Title: RNA-binding protein with serine-rich domain 1 regulates microsatellite instability of uterine corpus endometrial adenocarcinoma

doi: 10.6061/clinics/2021/e3318

Figure Lengend Snippet: (A) GSEA of the correlation between RNPS1 and Notch or Notch4 signaling pathway in UCEC. (B) Western blot assay for the analysis of the RNPS1, MSH1, MSH2, MSH6, and PMS2 expression levels in vivo . (B) Quantitation of RNPS1, MSH1, MSH2, MSH6, and PMS2 expression levels. (n=6, * p <0.05: sh-RNPS1 vs. Con; # p <0.05: IMR-1A+sh-RNPS1 vs. sh-RNPS1).

Article Snippet: The secondary antibodies used were anti-rabbit IgG (AS014) and anti-mouse IgG (H+L) (AS003), both from ABclonal (Wuhan, China), and IMR-1A (HY-100431A; MedChemExpress, Dallas, TX, USA).

Techniques: Western Blot, Expressing, In Vivo, Quantitation Assay

Journal: Nature Aging

Article Title: Modulating IL-11-dependent matrix stiffness to delay ovarian aging

doi: 10.1038/s43587-026-01159-2

Figure Lengend Snippet: a , Immunofluorescent staining of COL1A1 and TGFβ1. L and H represent regions with low and high coexpression of COL1A1 and TGFβ1, respectively. b , Measurement of ovarian matrix stiffness in COL1A1 lo TGFβ1 lo or COL1A1 hi TGFβ1 hi regions. Data are presented as the mean ± s.d. (Welch’s two-sided t -test). n = 20 per group. c , Schematic of RNA-seq of primary human ovarian fibroblasts (pHOFs) after in vitro TGFβ1 treatment (10 ng ml −1 , 24 h). d , IL11 RNA expression is upregulated in pHOFs stimulated with TGFβ1. Differential expression was assessed using a two-sided Wald test, with P values adjusted for multiple comparisons through the Benjamini–Hochberg method ( n = 3 per group). e , IL-11 protein abundance in human, mouse and rat ovaries across different age groups. f , Representative images of IL-11 immunohistochemical staining in ovaries from individuals of different ages or with ovarian dysfunction due to various diseases. g , Representative images of pHOFs immunostained for COL1A1 and ACTA2 after 24 h of incubation without stimulus (control), TGFβ1 (10 ng ml −1 ) or IL-11 (10 ng ml −1 ) or with TGFβ1 (10 ng ml −1 ) and an anti-IL-11 neutralizing antibody (2 μg ml −1 ). h , GO and KEGG enrichment analysis of differentially expressed genes in pHOFs incubated without stimulus (control) or IL-11 (10 ng ml −1 ) for 24 h. Red labels highlight GO terms enriched in pathways related to the ECM and its receptor. The analyses were performed using the hypergeometric test. P values were adjusted for multiple comparisons using the Benjamini–Hochberg FDR method. n = 3 per group. i , COL1A1 mRNA expression in pHOFs treated with IL-11 or IL-11 and NOTCH pathway inhibitor (PF-3084014, IMR-1), PI3K–AKT pathway inhibitor (PI3K/AKT-IN-1, BYL-719), cGMP–PKG pathway inhibitor (KT5823, MBP146-78), p38 MAPK pathway inhibitor (p38 MAPK-IN-1, SB 203580), ERK1/2 pathway inhibitor (ravoxertinib, SCH772984), JNK pathway inhibitor (JNK-IN-7, SP600125) or mTOR pathway inhibitor (rapamycin, everolimus). Red box highlighting indicates the ERK1/2 inhibitor group, which showed the most significant reduction in IL‑11‑induced COL1A1 upregulation. Data are presented as mean ± s.d., n = 3 for each group (unpaired two-tailed t -test or Welch’s two-tailed t -test). j , k , Human phospho-kinase array of pHOFs treated with 10 ng ml −1 IL-11 or vehicle control for 24 h. Numbers 1–8 on the blot ( j ) and the corresponding bars ( k ) indicate the following: 1) CREB at serine 133; 2) ERK1/2 at threonine 202 and tyrosine 204; 3) GSK3β at serine 9; 4) WNK1 at threonine 60; 5) β‑Catenin; 6) RSK1/2 at serine 221 and serine 227; 7) RSK1/2/3 at serine 380, serine 386 and serine 377; 8) STAT3 at tyrosine 705. Data are presented as the mean ± s.d., n = 3 for each group (unpaired two-tailed t -test). l , Western blot analysis of COL1A1 and ACTA2 expression in pHOFs treated with IL-11, with or without the ERK1/2 inhibitor SCH772984. m , Representative immunofluorescence images and quantification of COL1A1 and ACTA2 in pHOFs treated with IL-11 or IL-11 + SCH772984. int., integrated intensity. Data are presented as the mean ± s.d., n = 5 per group (one-way ANOVA).

Article Snippet: Experiments were carried out at low cell passages (≤ passage 3) and HOFs were treated with TGFβ1 (10 ng ml −1 ) or IL-11 (10 ng ml −1 ) in serum-free DMEM for 24 h. For pathway screening studies, HOFs were stimulated with IL-11 in the presence of PF-3084014 (10 μM, MedChemExpress), IMR-1 (10 μM, MedChemExpress), PI3K/AKT-IN-1 (5 μM, MedChemExpress), BYL-719 (250 nM, MedChemExpress), KT5823 (5 μM, MedChemExpress), MBP146-78 (150 nM, MedChemExpress), p38 MAPK-IN-1 (50 nM, MedChemExpress), SB 203580 (50 nM, MedChemExpress), ravoxertinib (5 nM, MedChemExpress), SCH772984 (5 nM, MedChemExpress), JNK-IN-7 (5 nM, MedChemExpress), SP600125 (100 nM, MedChemExpress), rapamycin (50 nM, MedChemExpress) or everolimus (10 nM, MedChemExpress) for 24 h. The anti-IL-11 antibody (R&D Systems, cat. no. MAB218, human) was used to neutralize IL-11 activity, and the Neutralization Dose (ND50) was 80 μg ml −1 in the presence of 10 ng ml −1 recombinant human IL-11.

Techniques: Staining, RNA Sequencing, In Vitro, RNA Expression, Quantitative Proteomics, Immunohistochemical staining, Incubation, Control, Expressing, Two Tailed Test, Western Blot, Immunofluorescence