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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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wnt inhibitors  (Tocris)
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HA-induced activation of the <t>canonical</t> <t>Wnt</t> <t>pathway</t> upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway <t>inhibitor</t> IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.
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HA-induced activation of the canonical Wnt pathway upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway inhibitor IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.

Journal: Frontiers in Immunology

Article Title: Hyaluronic acid−CD44 signaling from decidual stromal cells orchestrates dNK1 differentiation and immune tolerance in early pregnancy

doi: 10.3389/fimmu.2026.1777567

Figure Lengend Snippet: HA-induced activation of the canonical Wnt pathway upregulates FOSL2 to expand the dNK1-like subpopulation. (A) Analysis of transcriptional activity and expression levels of transcription factors in three dNK subsets using scRNA-seq data from normal decidual tissue ( n = 11). (B) Transcriptional activity and expression levels of FOSL2 in decidual tissues from normal pregnancy ( n = 5) and RSA groups ( n = 3). (C) Changes in NK92MI cells of the five transcription factors ( STAT3, MAFB, HES1, FOSL2, ETV5 ) characterized by high transcriptional activity and expression in the dNK1 subset following HMW-HA treatment, as determined by RT−qPCR ( n = 6 per group). (D) FOSL2 expression in NK92MI cells following HMW-HA treatment, assessed by WB ( n = 3 per group). (E) Wnt1 and β-catenin protein expression in NK92MI cells treated with or without HMW-HA in the presence or absence of CD44 blockade ( n = 3 per group). (F) FOSL2 expression in HA−treated NK92MI cells after addition of CD44−blocking antibody or the Wnt pathway inhibitor IWP−2 to the culture system ( n = 3 per group). Data are expressed as mean ± SD; * P < 0.05; ** P < 0.01; ns, not significant. EP, early pregnancy; SA, spontaneous abortion; RSA, recurrent spontaneous abortion.

Article Snippet: To determine the signaling pathways involved, we used an anti-CD44 blocking antibody (30 μg/mL; BioxCell, BE0262) to block CD44, and IWP-2 (50 μM; MedChemExpress, HY-13912) to inhibit the canonical Wnt pathway.

Techniques: Activation Assay, Activity Assay, Expressing, Quantitative RT-PCR, Blocking Assay