Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Differentially expressed genes in limbal epithelial stem/progenitor cell clusters compared to basal corneal epithelial cells isolated by laser capture microdissection (n = 5).

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Isolation, Laser Capture Microdissection, Standard Deviation, Binding Assay

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Differential expression of SOX family genes in limbal epithelial progenitor cell clusters compared to basal corneal epithelial cells isolated by laser capture microdissection (n = 5).

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Quantitative Proteomics, Isolation, Laser Capture Microdissection, Standard Deviation

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Immunohistochemical localization of SoxE family members in corneoscleral tissue sections. ( A ) Immunofluorescence microscopy demonstrates nuclear staining for Sox8 (clone 4E4.1) and Sox9 (clone 3C10) in suprabasal epithelial cells at the limbus (left column) and central cornea (right column), whereas Sox10 (clone BC34) is confined to few cells in the basal limbal epithelium. Higher magnification images of basal limbal regions (middle column), as indicated by boxed areas, show differential cytoplasmic (arrows) and nuclear localization of Sox8 and Sox9 in basal and suprabasal limbal epithelial cells. ( B ) High magnification images of individual channels show cytoplasmic localization of Sox9 in basal stem/progenitor cell clusters and nuclear localization in suprabasal limbal epithelial cells. ( C ) Double labeling experiments show nuclear co-localization of Sox8 (rabbit IgG) and Sox9 (clone 3C10) (left), distinct localization of Sox9 (clone 3C10) and Sox10 (rabbit IgG) (middle), and localization of Sox10 (clone BC34) to Melan A-positive cells (right). Nuclear counterstaining: DAPI. Individual channels of double labeling experiments are shown in Supplementary Fig. .

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Immunohistochemical staining, Immunofluorescence, Microscopy, Staining, Labeling

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Co-localisation of Sox9 with markers related to progenitor cell phenotype, differentiation and proliferation in the limbal epithelium. Double-labelling demonstrates co-localisation (arrows) of cytoplasmic Sox9 (red) with the stem/progenitor cell markers (green) N-cadherin, p75 nerve growth factor receptor (NGF-R), p63α, Oct4 and cytokeratin (CK) 15) in basal epithelial cells at the limbus. Suprabasal epithelial cells revealed co-localisation of nuclear Sox9 (red) with differentiation-related markers (green) CK3 and Pax6 as well as proliferation-related marker Ki-67. Sox9 monoclonal mouse antibody (clone 3C10) was used for double labelling experiments with polyclonal antibodies against Oct4, p63α, Pax6 and Ki-67, Sox9 polyclonal rabbit antibody (1) was used for double labelling experiments with monoclonal antibodies against p75 NGF-R, N-cadherin, CK3 and CK15. Nuclear counterstaining: DAPI. Individual channels of all double labelling experiments are shown in Supplementary Fig. .

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Marker, Bioprocessing

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Expression of Sox9 during limbal epithelial cell expansion and wound healing in vitro . ( A ) Relative expression levels of Sox9 in cultured limbal epithelial cells expanded as clones on a 3T3 feeder layer or as feeder-free monolayer in passage (P) 0 to P2. Expression levels were determined by quantitative real-time polymerase chain reaction (qRT-PCR) primer assays and normalized against GAPDH. Data are expressed as means (2 −∆CT × 1,000) ± SD (n = 3) relative to clonal cultures; *p < 0.01, unpaired t -test. ( B ) Limbal epithelial cell (LEPC) clones (dashed lines) on 3T3 feeder cells (3T3) stain positively for Sox9 (clone 3C10, red), preferentially towards the proliferating border of the clones; Sox9-expressing cells partly co-localize with Ki-67 (green). Nuclear staining: DAPI. ( C ) Immunofluorescent staining shows increased levels of nuclear Sox9 (clone 3C10) in basal/suprabasal limbal epithelial cells and central corneal epithelial cells after epithelial debridement and regeneration compared to unwounded control corneas (the background fluorescence seen in the central corneal stroma may be attributed to the epithelial debridement allowing media and serum components to infiltrate the stroma during the wound healing process). Higher magnification images of limbal epithelial progenitor cell clusters show increased nuclear localization of Sox9 in basal epithelial cells in wound healing conditions compared to cytoplasmic retention of Sox9 in control tissues (inserts). Nuclear staining: DAPI. ( D ) Relative expression of SOX9 in limbal epithelial cells of wounded and unwounded corneas as determined by quantitative real-time polymerase chain reaction (qRT-PCR) primer assays. Normalized data are expressed as means (2 −∆CT × 1,000) ± SD (n = 5) relative to unwounded controls; p = 0.08, unpaired t -test.

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Expressing, In Vitro, Cell Culture, Clone Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Staining, Control, Fluorescence

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Changes in mRNA expression following knockdown of SOX9 in cultured limbal epithelial cells by RNAi. ( A ) Results of quantitative real-time polymerase chain reaction (qRT-PCR) showing reduction of SOX9 mRNA transcripts in cultured limbal epithelial cells 24–96 hours after transfection with siRNA to SOX9 relative to control cells transfected with scramble siRNA (Control) (n = 6; Mean ± SD). ( B ) Transcriptional changes following knockdown of SOX9 in cultured limbal epithelial cells as determined by qRT-PCR. Significant or no relevant changes were seen in the expression levels of stemness-related genes ABCG2 (ATP Binding Cassette Subfamily G Member 2), TP63 (ΔNp63α) and CEBPD (CCAAT/enhancer-binding protein delta); progenitor cell marker genes KRT15 (keratin 15), KRT14 and CDH2 (N-cadherin); differentiation marker genes KRT3, KRT12 and IVL (involucrin); and proliferation-related genes PCNA (proliferating cell nuclear antigen), CDKN1A (cyclin-dependent kinase inhibitor 1A, p21) and CDKN1C (p57). Normalized data are expressed as means (2 −∆CT × 1,000) ± SD (n = 6) relative to scramble siRNA-transfected control cells; *p < 0.05, **p < 0.01, ***p < 0.001, unpaired t -test.

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Expressing, Knockdown, Cell Culture, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Control, Binding Assay, Marker

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Changes in protein expression and proliferation following knockdown of SOX9 in cultured limbal epithelial cells by RNAi. ( A ) Western blot analysis of limbal epithelial cells transfected with either siRNA that targets SOX9 or non-targeting, scrambled siRNA as a control. Protein expression of Sox9, cytokeratin (CK) 15, CK3 and PCNA was detected with monoclonal antibodies, normalized to the house-keeping gene ß-actin, and expressed as percent of the expression in control cells (Ctrl); (n = 3; mean ± SD); *p < 0.05, **p < 0.005, ***p < 0.0001, unpaired t -test. Uncropped versions of Western blots are shown in Supplementary Fig. . ( B ) BrdU incorporation (i.e., cell proliferation) was determined by measuring absorbance at 450 nm. Statistically significant differences were observed at 72 (*p = 0.005) and 96 hours (**p = 0.009) between cells transfected with siRNA that targets SOX9 (Si-Sox9) and control cells transfected with scramble siRNA (Scr-Crtl) (n = 3; mean ± SD).

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Expressing, Knockdown, Cell Culture, Western Blot, Transfection, Control, Bioprocessing, BrdU Incorporation Assay

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Interactions between Sox9 and cell signaling pathways. ( A ) Changes in mRNA expression of genes centrally involved in Wnt/ß-catenin signaling, i.e., WNT4 (Wnt-4), CTNNB1 (ß-catenin) and GSK3B (glycogen synthase kinase 3 beta), following knockdown of SOX9 in cultured limbal epithelial cells by RNAi relative to mock-transfected control cells. Normalized data are expressed as means (2 −∆CT × 1,000) ± SD (n = 3); *p < 0.05, **p < 0.01, ***p < 0.001, unpaired t -test. ( B ) Changes in SOX9 mRNA expression following exposure of cultured limbal epithelial cells with signaling activators lithium chloride (LiCl), IM-12, BMP-2, JAG-1 and SAG as well as signaling inhibitors C-59, DMH1, DAPT and Cyclopamine (Cyclo) for 24 hours relative to vehicle-treated control cells (Ctrl). Normalized data are expressed as means (2 −∆CT × 1,000) ± SD (n = 3); *p < 0.05, **p < 0.01, unpaired t -test. ( C ) Changes in Sox9 protein expression following exposure of cultured limbal epithelial cells to Hedgehog signaling activators Sonic hedgehog (SHH), SAG and Purmorphamine (Purmo); BMP-2; Wnt signaling activators Wnt-3a, lithium chloride (LiCl) and IM-12; and Notch signaling ligand JAG-1 for 48 hours relative to vehicle-treated control cells (Ctrl). Sox9 protein expression was detected with the monoclonal antibody (clone 3C10), normalized to the house-keeping gene ß-actin, and expressed as percent of the expression in control cells (Ctrl); (n = 3; mean ± SD). Uncropped versions of Western blots are shown in Supplementary Fig. .

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Protein-Protein interactions, Expressing, Knockdown, Cell Culture, Transfection, Control, Western Blot

Journal: Scientific Reports

Article Title: Transcription factor profiling identifies Sox9 as regulator of proliferation and differentiation in corneal epithelial stem/progenitor cells

doi: 10.1038/s41598-018-28596-3

Figure Lengend Snippet: Model illustrating the mutually repressive interaction of Sox9 and Wnt/ß-catenin signaling as well as potentially involved regulatory signaling pathways in the limbal stem cell niche (mod. after Xu, Z. et al ., Elife 4, e10567 (2015) ( https://creativecommons.org/licenses/by/4.0 ).

Article Snippet: Membranes were blocked with SuperBlock T20 Blocking Buffer (Thermo Fisher Scientific) for 1 hour and incubated overnight using monoclonal mouse antibodies against Sox9 (1:5000; clone 3C10; BioRad), Cytokeratin 3/76 (1:5000; clone AE5; Millipore), Cytokeratin 15 (1:1000; clone EPR1614Y; Abcam) and PCNA (1:5000; clone PC10; Abcam).

Techniques: Protein-Protein interactions

Journal: Journal of Orthopaedic Translation

Article Title: HIF-2α/TFR1 mediated iron homeostasis disruption aggravates cartilage endplate degeneration through ferroptotic damage and mtDNA release: A new mechanism of intervertebral disc degeneration

doi: 10.1016/j.jot.2024.03.005

Figure Lengend Snippet: TfR1 knockdown restored oxidative stress and TNF-α induced CEP degeneration. A Representative immunofluorescence images of COL2 in TBHP and TNF-α treated CEP chondrocytes with or without TfR1 siRNA transfection. Scale bar = 50 μm. B–C Representative western blotting images of TfR1, MMP3, MMP13, SOX9, COL2 and semi-quantitative analysis of band density in TBHP treated CEP chondrocytes. D-E Representative western blotting images of COL10, RUNX2 and semi-quantitative analysis of band density in TBHP treated CEP chondrocytes. F Alizarin Red staining for calcium deposition in endplate chondrocytes. G Semi-quantitative analysis of the mineralized nodule in endplate chondrocytes. H–I Annexin V-FITC/PI flow cytometric analysis was conducted to evaluate apoptosis rate induced by TBHP and TfR1 siRNA transfection. J-K Representative western blotting images of TfR1, MMP3, MMP13, SOX9, COL2 and semi-quantitative analysis of band density in TNF-α treated CEP chondrocytes. Data are shown as the mean ± SD of at least three independent experiments. The P value is indicated by stars: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The primary antibodies used were as follows: TfR1 (#ab84036, Abcam, dilution 1:1000), HIF-2α(#BM3997, Boster, dilution 1:1000), IRF3(#4302, Cell Signaling Technology, dilution 1:1000), p-IRF3(Ser396, #29047, Cell Signaling Technology, dilution 1:1000), BNIP3(#3769S, Cell Signaling Technology, dilution 1:1000), GPX4 (#ab125066, Abcam, dilution 1:5000), SLC7A11 (#26864-1-AP, Proteintech, dilution 1:1000), cGAS (#A8335, ABclonal, dilution 1:1000), STING (#19851-1-AP, Proteintech, dilution 1:1000), DRP1 (#12957-1-AP, Proteintech, dilution 1:1000), MFF(#17090-1-AP, Proteintech, dilution 1:1000), FIS1(#10956-1-AP, Proteintech, dilution 1:1000), MMP3(#BM4074, Boster, dilution 1:1000), MMP13(#18165-1-AP, Proteintech, dilution 1:1000), COL2 (#28459-1-AP, Proteintech, dilution 1:1000), SOX9 (#A00177-2, Boster, dilution 1:1000), COL10 (#BA2023, Boster, dilution 1:1000), RUNX2 (#PB0171, Boster, dilution 1:1000), GAPDH (#10491-1-AP, Proteintech, dilution 1:5000).

Techniques: Knockdown, Immunofluorescence, Transfection, Western Blot, Staining

Journal: Journal of Orthopaedic Translation

Article Title: HIF-2α/TFR1 mediated iron homeostasis disruption aggravates cartilage endplate degeneration through ferroptotic damage and mtDNA release: A new mechanism of intervertebral disc degeneration

doi: 10.1016/j.jot.2024.03.005

Figure Lengend Snippet: TfR1 inhibitor ameliorated CEP calcification and IDD development. A Eight-week aged mice were randomly divided in three groups and intra-peritoneally injected Feristatin II solution followed by LSI surgery. HE staining of L4/5 intervertebral discs in the Ctrl group, IDD group, and Fer-II group. scale bar = 200 μM. B Histological score of the L4–5 segments of the lumbar spine among the three groups and the ratio of positive cells for TFR1, SOX9, MMP3 from each group. C-E Immunohistochemistry for TfR1, MMP3, SOX9 in the Ctrl, IDD, and Fer-II groups. Scale bar = 100 μm. F Micro-CT analysis, 3D reconstruction, and X-ray images of cartilage endplate. G Quantification of cartilage endplate calcification via microarchitecture parameters [bone volume per tissue volume (BV/TV)]. Data are shown as the mean ± SD. N = 5, The P value is indicated by stars: ***P < 0.001, ****P < 0.0001.

Article Snippet: The primary antibodies used were as follows: TfR1 (#ab84036, Abcam, dilution 1:1000), HIF-2α(#BM3997, Boster, dilution 1:1000), IRF3(#4302, Cell Signaling Technology, dilution 1:1000), p-IRF3(Ser396, #29047, Cell Signaling Technology, dilution 1:1000), BNIP3(#3769S, Cell Signaling Technology, dilution 1:1000), GPX4 (#ab125066, Abcam, dilution 1:5000), SLC7A11 (#26864-1-AP, Proteintech, dilution 1:1000), cGAS (#A8335, ABclonal, dilution 1:1000), STING (#19851-1-AP, Proteintech, dilution 1:1000), DRP1 (#12957-1-AP, Proteintech, dilution 1:1000), MFF(#17090-1-AP, Proteintech, dilution 1:1000), FIS1(#10956-1-AP, Proteintech, dilution 1:1000), MMP3(#BM4074, Boster, dilution 1:1000), MMP13(#18165-1-AP, Proteintech, dilution 1:1000), COL2 (#28459-1-AP, Proteintech, dilution 1:1000), SOX9 (#A00177-2, Boster, dilution 1:1000), COL10 (#BA2023, Boster, dilution 1:1000), RUNX2 (#PB0171, Boster, dilution 1:1000), GAPDH (#10491-1-AP, Proteintech, dilution 1:5000).

Techniques: Injection, Staining, Immunohistochemistry, Micro-CT

Journal: Journal of Orthopaedic Translation

Article Title: HIF-2α/TFR1 mediated iron homeostasis disruption aggravates cartilage endplate degeneration through ferroptotic damage and mtDNA release: A new mechanism of intervertebral disc degeneration

doi: 10.1016/j.jot.2024.03.005

Figure Lengend Snippet: TfR1 mediated mtDNA release plays essential roles in the cGAS-STING activation and CEP chondrocytes degeneration. A Representative western blotting images of cGAS, STING, p-IRF3, IRF3 and semi-quantitative analysis of band density in TBHP treated CEP chondrocytes with or without TfR1 siRNA transfection. B Mitochondrial membrane potential (MMP) was detected by fluorescence after chondrocytes were incubated with JC-1. Red fluorescence represents JC-1 aggregates in healthy mitochondria, while green fluorescence is emitted by JC-1 monomers, representing MMP dissipation. Merged images exhibit co-localization of JC- 1 aggregates and monomers. C Representative fluorescence images of dsDNA (green) and mitochondria (red) in the TBHP and TNF-α treated CEP chondrocytes with or without TfR1 siRNA transfection. scale bars = 50 μm. D CEP chondrocytes were pretreated with ethidium bromide 48h, then 100 μM TBHP was added and western blot was conducted to examine cGAS, STING, p-IRF3, IRF3 proteins expression. The band density was quantified and normalized to control. E Representative fluorescence images of dsDNA (green) and mitochondria (red) in the TBHP and EthBr treated CEP chondrocytes. F Representative western blotting images of SOX9, COL2, MMP3, MMP13 and semi-quantitative analysis of band density in TBHP and EthBr treated CEP chondrocytes. Data are shown as the mean ± SD of at least three independent experiments. The P value is indicated by stars: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The primary antibodies used were as follows: TfR1 (#ab84036, Abcam, dilution 1:1000), HIF-2α(#BM3997, Boster, dilution 1:1000), IRF3(#4302, Cell Signaling Technology, dilution 1:1000), p-IRF3(Ser396, #29047, Cell Signaling Technology, dilution 1:1000), BNIP3(#3769S, Cell Signaling Technology, dilution 1:1000), GPX4 (#ab125066, Abcam, dilution 1:5000), SLC7A11 (#26864-1-AP, Proteintech, dilution 1:1000), cGAS (#A8335, ABclonal, dilution 1:1000), STING (#19851-1-AP, Proteintech, dilution 1:1000), DRP1 (#12957-1-AP, Proteintech, dilution 1:1000), MFF(#17090-1-AP, Proteintech, dilution 1:1000), FIS1(#10956-1-AP, Proteintech, dilution 1:1000), MMP3(#BM4074, Boster, dilution 1:1000), MMP13(#18165-1-AP, Proteintech, dilution 1:1000), COL2 (#28459-1-AP, Proteintech, dilution 1:1000), SOX9 (#A00177-2, Boster, dilution 1:1000), COL10 (#BA2023, Boster, dilution 1:1000), RUNX2 (#PB0171, Boster, dilution 1:1000), GAPDH (#10491-1-AP, Proteintech, dilution 1:5000).

Techniques: Activation Assay, Western Blot, Transfection, Membrane, Fluorescence, Incubation, Expressing, Control

Journal: The ocular surface

Article Title: Immuno Tomography (IT) and Imaging Mass Cytometry (IMC) for constructing spatially resolved, multiplexed 3D IMC data sets

doi: 10.1016/j.jtos.2022.04.008

Figure Lengend Snippet: Antibodies used for IMC.

Article Snippet: Sm147 , Sox9 , Fluidigm , EPR14335 , 3147022D , 1:50 , 10.

Techniques: Concentration Assay

Journal: Experimental & Molecular Medicine

Article Title: (Non)canonical Wnt signaling, cytoarchitecture and stemness: new insights from primary nonmetastatic, primary metastatic, regional and distant metastatic models of adrenocortical carcinoma

doi: 10.1038/s12276-025-01507-z

Figure Lengend Snippet: a Immunofluorescence for KLF4, Oct4, Nanog, Sox2, c-Myc, CD24, CD44, CD68, Nestin, ALDH1A1, ALDH1A3, Foxa1 and Sox9 in TVBF-7, NCI-H295R and MUC-1 spheroids, presented as single stainings together with DAPI in each model. Scale bar, 50 μm. b Gene expression analysis of the various stem cell markers under control and treated conditions in monolayer cultures from all three cell models. Drug treatments from left to right: control (C), forskolin (F), mitotane (M), JQ1 (J) and THZ-1 (T).

Article Snippet: SOX9 , CSB-RA202969A0HU, Cusabio (Houston, TX, USA) , 1/200.

Techniques: Immunofluorescence, Gene Expression, Control