Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Analysis of PHF6 expression across cancer types. ( A ) PHF6 expression levels in different types of cancer in the TCGA database. ( B ) PHF6 expression levels in combination with the GTEx and TCGA databases. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ( C – F ) IHC images of PHF6 expression in normal lung ( C ), lung cancer ( D ), prostate ( E ), and prostate cancer ( F ) from the HPA database . Magnification: 20 × .

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Expressing

Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Prognostic significance of PHF6 in the TCGA dataset. ( A ) The association between PHF6 expression and OS. ( B ) The association between PHF6 expression and DSS. ( C , D ) The association between PHF6 and OS in PAAD ( C ) and GBMLGG cohorts ( D ). ( E , F ) The association between PHF6 and DSS in LIHC ( E ) and KIRC cohorts ( F ).

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Expressing

Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Tumor immune analysis in PHF6 expression. The correlations between PHF6 with stromal score ( A – D ), immune score ( E – H ), and ESTIMATE score ( I – L ) in GBMLGG, LGG, SARC, and LUSC. The correlation of PHF6 with immune infiltration levels based on CIBERSORT algorithm ( M ). * p < 0.05.

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Expressing

Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Enrichment analysis and drug sensitivity analysis of PHF6. GO analysis including biological processes ( A ), cellular components ( B ), and molecular functions ( C ) showed that PHF6 was enriched in cell cycle and chromatin-binding pathways ( A – C ). KEGG analysis indicated that PHF6 was related to spliceosome pathways ( D ). Impact of PHF6 on drug sensitivity in LIHC ( E – H ) and PAAD ( I – L ).

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Binding Assay

Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Correlations of PHF6 and TMB ( A ), microsatellite instability ( B ), DNAss ( C ), and RNAss ( D ). Correlations between PHF6 expression and RNA m1A modification ( E ). * p < 0.05.

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Expressing, Modification

Journal: Diagnostics

Article Title: Systematic Pan-Cancer Analysis Identifies PHF6 as an Immunological and Prognostic Biomarker

doi: 10.3390/diagnostics16010110

Figure Lengend Snippet: Knockout of PHF6 suppressed cell proliferation of liver cancer and PAAD cells. Western blot detected the knockout efficiency of PHF6 ( A ). PHF6 antibody (Santa Cruz Biotechnology, Inc., Dallas, TX, United States), β-tubulin (ZSGB-BIO, Beijing, China). sgC: sgControl, sg1: sgPHF6#, sg2: sgPHF6#2. The CCK-8 experiment on the proliferation of HepG2 ( B ), Huh7 ( C ), CFPAC-1 ( D ), and Mia-Paca-1 ( E ). Effect of PHF6 knockout colony formation ( F ) of HepG2 ( G ), Huh7 ( H ), CFPAC-1 ( I ), and Mia-Paca-1 ( J ). ** p < 0.01, *** p < 0.001, **** p < 0.0001, ***** p < 0.00001, ****** p < 0.000001, ******* p < 0.0000001, ******** p < 0.00000001.

Article Snippet: Immunohistochemistry results for PHF6 in both cancerous and normal tissues were retrieved from the Human Protein Atlas (HPA) database [ ].

Techniques: Knock-Out, Western Blot, CCK-8 Assay

Journal: Molecular Psychiatry

Article Title: Regulation of stress susceptibility by chromatin-binding protein PHF6 in the pituitary intermediate lobe

doi: 10.1038/s41380-025-03300-w

Figure Lengend Snippet: a Schematic illustration of the generation of Phf6-CreER knock-in (KI) mice and the experimental strategy for labeling IL PHF6 cells through stereotactic injection of AAV-DIO-mCherry into the pituitary intermediate lobe of adult male Phf6-CreER mice, followed by tamoxifen-induced recombination. b Representative images show colocalization of endogenous PHF6 (green) and mCherry (red) in the pituitary intermediate lobe. Scale bars, 100 μm. c Quantification revealed that 97.5% ± 0.3% of mCherry-positive cells co-expressed endogenous PHF6 ( n = 3). d Experimental design for IL PHF6 cell inhibition and subsequent evaluation of stress-induced anxiety-like behaviors. e Left: Representative images showing expression of hM4D-mCherry (or mCherry) in the IL of Phf6-CreER mice, co-stained with c-Fos (green). Right: Quantification demonstrated significantly reduced c-Fos expression in hM4D-expressing IL cells compared to mCherry controls after RS-stress (mCherry, n = 3; hM4D, n = 3, *** p < 0.001, unpaired Student’s t -test). DAPI is shown in blue. Scale bars, 50 μm. f Representative locomotor trajectories and quantification of time spent in the center area of the OF test. Inhibition of IL PHF6 cells significantly increased the time spent in the center area of the OF test following RS stress (mCherry-saline, n = 9; hM4D-saline, n = 10; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, *** p < 0.001, two-way ANOVA with Bonferroni’s multiple comparisons test). g Representative locomotor trajectories and quantification of time spent in the open arms of the EPM test. Inhibition of IL PHF6 cells significantly increased the time spent in the open arms of the EPM test after RS stress (mCherry-saline, n = 9; hM4D-saline, n = 9; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, ** p < 0.01, two-way ANOVA with Bonferroni’s multiple comparisons test). h Quantification of the latency to feed in the NSF test. Inhibition of IL PHF6 cells significantly decreased the latency to feed in the NSF test following RS stress (mCherry-saline, n = 9; hM4D-saline, n = 10; mCherry-CNO: n = 9; hM4D-CNO: n = 10; * p < 0.05, ** p < 0.01, two-way ANOVA with Bonferroni’s multiple comparisons test. Data are presented as mean ± SEM.

Article Snippet: For chemogenetic and circuit tracing experiments, Phf6-CreER mice (C57BL/6J background, generated by Cyagen Biosciences) were used.

Techniques: Knock-In, Labeling, Injection, Inhibition, Expressing, Staining, Saline

Journal: Molecular Psychiatry

Article Title: Regulation of stress susceptibility by chromatin-binding protein PHF6 in the pituitary intermediate lobe

doi: 10.1038/s41380-025-03300-w

Figure Lengend Snippet: a Experimental design for IL PHF6 cell activation and assessment of anxiety-like behaviors. b Left: Representative images showing hM3D-mChery (or mCherry) expression in the IL of Phf6-CreER mice, co-stained with c-Fos (green). Right: Quantification revealed significantly higher c-Fos expression in hM3D-expressing IL PHF6 cells compared to mCherry controls (mCherry, n = 3; hM3D, n = 3, **** p < 0.0001, unpaired Student’s t -test). DAPI is shown in blue. Scale bars, 50 μm. c Representative locomotor trajectories and quantification of time spent in the center area of the OF test. Activation of IL PHF6 cells significantly decreased the time spent in the center area of the OF test (mCherry-saline, n = 8; hM3D-saline, n = 8; mCherry-CNO, n = 10; hM3D-CNO, n = 8; * p < 0.05, two-way ANOVA with Bonferroni’s multiple comparisons test). d Representative locomotor trajectories and time spent in the open arms of the EPM test. Activation of IL PHF6 cells significantly decreased the time spent in the open arms of the EPM test (mCherry-saline, n = 8; hM3D-saline, n = 8; mCherry-CNO, n = 10; hM3D-CNO, n = 8; * p < 0.05, two-way ANOVA with Bonferroni’s multiple comparisons test). e Quantification of the latency to feed in the NSF test. Activation of IL PHF6 cells significantly increased the latency to feed in the NSF test (mCherry-saline, n = 8; hM3D-saline, n = 8; mCherry-CNO, n = 10; hM3D-CNO, n = 8; * p < 0.05, two-way ANOVA with Bonferroni’s multiple comparisons test). f Experimental design for assessing hormone release following IL PHF6 cell activation. g-i Plasma quantification showed significantly elevated α-MSH ( g ), β-endorphin ( h ), and corticosterone ( i ) levels in hM3D-expressing mice compared to controls. (mCherry, n = 10; hM3D, n = 8, * p < 0.05, unpaired Student’s t -test). j Schematic diagram of retrograde monosynaptic tracing using rabies virus in Phf6-CreER mice. Starter cells are labeled in yellow, as indicated by the white arrowheads. Scale bars, 100 μm. k Rabies-labeled upstream inputs to IL PHF6 cells. Scale bars, 100 μm. l Percentage of RV-traced tdTomato-positive neurons located in the paraventricular nucleus (PVN), the periventricular zone (Pe), and the arcuate nucleus (ARC) ( n = 3). m Representative images of RV-traced tdTomato-positive neurons co-expressing CRH (green) in the PVN. Scale bars, 100 μm (low magnification) and 10 μm (high magnification). Data are presented as mean ± SEM.

Article Snippet: For chemogenetic and circuit tracing experiments, Phf6-CreER mice (C57BL/6J background, generated by Cyagen Biosciences) were used.

Techniques: Activation Assay, Expressing, Staining, Saline, Clinical Proteomics, Virus, Labeling