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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene <t>NR2F2.</t> (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.
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Validation of model genes and preliminary exploration of core gene NR2F2. (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.

Journal: Frontiers in Immunology

Article Title: NR2F2 in cancer-associated fibroblasts drives immune microenvironment remodeling and promotes lung adenocarcinoma progression

doi: 10.3389/fimmu.2026.1776008

Figure Lengend Snippet: Validation of model genes and preliminary exploration of core gene NR2F2. (A) Transcriptional levels of HGF, PEBP1, TIMP1, NR2F2, and SHC1 in LUAD tissues relative to surrounding non-cancerous tissues were validated using RT-qPCR. (B) Successful induction was confirmed by Western blot analysis of CAF activation markers. (C) Differential expression of HGF, PEBP1, TIMP1, NR2F2, and SHC1 between CAFs and NAFs was detected by RT-qPCR. (D) Basal expression levels of NR2F2 in CAFs and NAFs were measured by Western blot. (E) Comparative Western blot results of NR2F2 expression in LUAD tissues versus surrounding healthy tissues. (F) Representative IHC staining images of NR2F2 in TMA, tissue microarray cohort (Scale bars: upper panel, 200 µm; lower panel, 50 µm). (G) Immunofluorescence staining images of α-SMA (green), NR2F2 (red), and DAPI (blue) in lung adenocarcinoma tissue sections (scale: 25 µm). (H) Dot plot analysis of NR2F2 expression levels across different cell types. (I) GSEA enrichment analysis revealed signal-enriched pathways of NR2F2. n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.

Article Snippet: The NR2F2 overexpression plasmid was purchased from Vigene Biosciences (China), and the empty pEnter vector was used as a negative control.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Western Blot, Activation Assay, Quantitative Proteomics, Expressing, Immunohistochemistry, Microarray, Immunofluorescence, Staining

Co-culture with NR2F2 overexpression CAFs promote the proliferation, migration and invasion ability of tumor cells. (A) Expression levels of α-SMA, FAP, and NR2F2 proteins in NAFs, normal-associated fibroblasts and CAFs,cancer-associated fibroblasts were analyzed by Western blotting and quantitative detection under specific conditions. (B) Relative mRNA expression levels of HGF, IL-6, FGF2, and PDGFD were detected by RT-qPCR in CAF-Vec and CAF-OE groups. (C) Schematic diagram of co-culture system between CAFs and tumor cells. (D) Proliferation efficiency was validated by CCK8 assay. (E) Proliferation efficiency was evaluated by colony formation assay. (F) Proliferation efficiency was confirmed by EdU assay (scale: 100 µm). (G) Proliferation efficiency was assessed by wound healing experiment (scale: 400 µm). (H) Migration and invasion capabilities were verified by Transwell assay (scale: 200 µm). n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.

Journal: Frontiers in Immunology

Article Title: NR2F2 in cancer-associated fibroblasts drives immune microenvironment remodeling and promotes lung adenocarcinoma progression

doi: 10.3389/fimmu.2026.1776008

Figure Lengend Snippet: Co-culture with NR2F2 overexpression CAFs promote the proliferation, migration and invasion ability of tumor cells. (A) Expression levels of α-SMA, FAP, and NR2F2 proteins in NAFs, normal-associated fibroblasts and CAFs,cancer-associated fibroblasts were analyzed by Western blotting and quantitative detection under specific conditions. (B) Relative mRNA expression levels of HGF, IL-6, FGF2, and PDGFD were detected by RT-qPCR in CAF-Vec and CAF-OE groups. (C) Schematic diagram of co-culture system between CAFs and tumor cells. (D) Proliferation efficiency was validated by CCK8 assay. (E) Proliferation efficiency was evaluated by colony formation assay. (F) Proliferation efficiency was confirmed by EdU assay (scale: 100 µm). (G) Proliferation efficiency was assessed by wound healing experiment (scale: 400 µm). (H) Migration and invasion capabilities were verified by Transwell assay (scale: 200 µm). n ≥ 3, **p < 0.01, ***p < 0.001, **** p < 0.0001.

Article Snippet: The NR2F2 overexpression plasmid was purchased from Vigene Biosciences (China), and the empty pEnter vector was used as a negative control.

Techniques: Co-Culture Assay, Over Expression, Migration, Expressing, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, EdU Assay, Transwell Assay