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fluoromount g with dapi  (SouthernBiotech)


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    Structured Review

    SouthernBiotech fluoromount g with dapi
    Fluoromount G With Dapi, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 97/100, based on 10471 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dapi+fluoromount+g/Fluoromount-G/pm41746362-309-4-7
    Average 97 stars, based on 10471 article reviews
    fluoromount g with dapi - by Bioz Stars, 2026-10
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    Related Articles

    Microscopy:

    Article Title: Revealing abrupt transitions from goal-directed to habitual behavior.
    Article Snippet: The next day, the slices were washed with 0.2% PBST at RT on shaker for 3 times, 5min each, then stained with secondary antibody (Alexa Fluor® 488 AffiniPureTM Donkey Anti-Goat IgG, 1:500 dilution in 0.2% PBST, Jackson Immuno #705-545- 003, ex488nm/em496nm) at RT on shaker for 2h. .. The slices were washed with 1XPBS at RT on a shaker for 3 times, 5min each, before mounted on microscope slides (Premium Superfrost® Plus Microscope Slides, VWR # 48311-703) with DAPI Fluoromount-G® (SouthernBiotech #0100-20, ex405nm/em465nm), and imaged with Zeiss LSM 700 confocal microscope under 10X magnification. .. All analyses were performed using custom-written MATLAB code (The MathWorks, 2019b, 2021a, or 2022a) or R environment.

    Article Title: Methods of gene therapy
    Article Snippet: After washing 3×5 minutes in PBS, sections were incubated in secondary antibody (Goat anti-Rabbit Alexa Fluor® 555, Invitrogen cat. A27039 (Carlsbad, CA), diluted 1/2000) for 1 hour at room temperature. .. Sections were mounted in DAPI Fluoromount-G® (Southern Biotech, Inc., Birmingham, AL) and visualized using a Zeiss confocal microscope. ..

    Staining:

    Article Title: SPT6 maintains epidermal homeostasis by inhibiting an NF-κB-positive feedback loop to prevent excessive inflammation.
    Article Snippet: The secondary antibodies used were Alexa 594-conjugated donkey anti-mouse IgG (Invitrogen: A21203), Alexa 488-conjugated goat anti-rabbit IgG (Invitrogen: A11034) and Alexa 594-conjugated donkey anti-rat IgG (Invitrogen: A21209) at 1:500. .. Nuclei were stained with DAPI Fluoromount-G (Southern Biotech: 0100-20). .. Images were acquired using an Olympus/BX63 upright fluorescence microscope.

    Article Title: Immunomodulatory hydrogel reprograms IL-17/NF-κB signaling to drive regeneration in diabetic wounds
    Article Snippet: After washing with PBS, tissues were treated with Alexa Fluor 647-conjugated goat anti-rabbit IgG (1:1000; Abcam, UK) and Alexa Fluor 488-conjugated goat anti-rat IgG (1:1000; Abcam, UK) for 2 h in a humidified chamber. .. Nuclei were stained with DAPI fluoromount-g® (Southern Biotech, USA). .. Images were captured using a fluorescence microscope (Carl Zeiss, Germany) and analyzed with ImageJ software.

    Incubation:

    Article Title: PACS1 syndrome mutation disrupts dynein-mediated cargo transport via HDAC6 and BICD2
    Article Snippet: Cells grown on coverslips were fixed with 4% (w/v) paraformaldehyde at RT for 20 min, permeabilized, and blocked with PBS containing 0.5% Triton X-100 and 2% normal goat serum (Gibco, PCN5000) for 15 min. .. Samples were then incubated with primary antibodies for 2 h at RT or overnight at 4 °C, washed 3 times with PBS, and incubated with fluorophore-conjugated secondary antibodies at RT for 1 h. Finally, coverslips were mounted with DAPI-Fluoromount G (Southern Biotech, 0100-20) and incubated overnight in the dark. .. Images were captured using a Nikon Ti2-E confocal microscope equipped with a resonant scanner and processed with the Nikon Elements analytical software.



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    (A) Experimental paradigm for spontaneous withdrawal consisting of 7 days of twice daily escalating doses of morphine (or saline control), followed by a single final dose and 24 hours of abstinence before tissue collection. (B) Example images of anti-Fos immunostaining after spontaneous withdrawal paradigm, counterstained with <t>DAPI.</t> Dotted lines indicate boundaries of MHb and LHb. Arrow indicates Fos + cluster in the LHb. (C) Average distribution of Fos + cells across anterior-posterior axis of the habenula for each treatment condition. Contour lines indicate density of Fos + cells, normalized to the maximum density in the saline condition. (D) Quantification of the density of Fos + neurons in saline control and spontaneous withdrawal conditions. Each circle represents the mean for an individual animal. n=186 sections total from 10 mice per condition. **p< 0.01 by t-test. (E) Quantification of Fos + cells in all sections, binned every 400 μm along the A-P axis. Each circle represents one tissue section. (F) Same data in D, separated by sex. Two-Way ANOVA, treatment effect, **p< 0.01; sex effect, n.s.; interaction, n.s. Fisher’s post hoc, **p< 0.01. Open circles indicate female, filled circles indicate male.
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    (A) Experimental paradigm for spontaneous withdrawal consisting of 7 days of twice daily escalating doses of morphine (or saline control), followed by a single final dose and 24 hours of abstinence before tissue collection. (B) Example images of anti-Fos immunostaining after spontaneous withdrawal paradigm, counterstained with <t>DAPI.</t> Dotted lines indicate boundaries of MHb and LHb. Arrow indicates Fos + cluster in the LHb. (C) Average distribution of Fos + cells across anterior-posterior axis of the habenula for each treatment condition. Contour lines indicate density of Fos + cells, normalized to the maximum density in the saline condition. (D) Quantification of the density of Fos + neurons in saline control and spontaneous withdrawal conditions. Each circle represents the mean for an individual animal. n=186 sections total from 10 mice per condition. **p< 0.01 by t-test. (E) Quantification of Fos + cells in all sections, binned every 400 μm along the A-P axis. Each circle represents one tissue section. (F) Same data in D, separated by sex. Two-Way ANOVA, treatment effect, **p< 0.01; sex effect, n.s.; interaction, n.s. Fisher’s post hoc, **p< 0.01. Open circles indicate female, filled circles indicate male.
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    (A) Immgen ULI RNA sequencing data showing expression of Mrgpra6 in a subset of immune cells. (B) Bulk RNA sequencing of sorted lung basophils showing normalized transcripts per million (TPM) expression of highly expressed basophil genes Cd200R3, Mcpt8, and Il4 compared to Mrgpra6 (n = 5). (C) Flow cytometry gating of live, CD45 + blood cells identifying the relative contribution of basophils, eosinophils, neutrophils, and mast cell progenitors to the tdTomato + population. (D) Representative confocal images of sections of the lung from Mrgpra6CreER; Rosa26LSL-tdTomato mice showing <t>DAPI</t> (blue) and tdTomato (red) expression in Mrgpra6-expressing cells. (E-F) Flow cytometric analysis of lung basophils demonstrates that ∼80% are tdTomato + , consistent with Mrgpra6 expression in this population (E). Conversely, ∼80% of lung tdTomato+ cells are Cd200r3+ FcerI+ basophils, supporting the specificity of Mrgpra6 to basophils in the lung (F) (n = 9).
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    (A) Immgen ULI RNA sequencing data showing expression of Mrgpra6 in a subset of immune cells. (B) Bulk RNA sequencing of sorted lung basophils showing normalized transcripts per million (TPM) expression of highly expressed basophil genes Cd200R3, Mcpt8, and Il4 compared to Mrgpra6 (n = 5). (C) Flow cytometry gating of live, CD45 + blood cells identifying the relative contribution of basophils, eosinophils, neutrophils, and mast cell progenitors to the tdTomato + population. (D) Representative confocal images of sections of the lung from Mrgpra6CreER; Rosa26LSL-tdTomato mice showing <t>DAPI</t> (blue) and tdTomato (red) expression in Mrgpra6-expressing cells. (E-F) Flow cytometric analysis of lung basophils demonstrates that ∼80% are tdTomato + , consistent with Mrgpra6 expression in this population (E). Conversely, ∼80% of lung tdTomato+ cells are Cd200r3+ FcerI+ basophils, supporting the specificity of Mrgpra6 to basophils in the lung (F) (n = 9).
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    Ferritinophagy aggravated MV-induced pulmonary fibrosis through regulating ferroptosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ferritinophagy markers NCOA4 and FTH in MLE-12 cells with and without MS. n = 3 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. (G, H). Immunoblot and quantitative analysis showing the expression levels of SLC7A11, GPX4 and FTH in DFO-treated MLE-12 cells with and without MS. n = 3 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (I), or FTH (Green) and NCOA4 (Red) in the lung tissues (J). White arrows point to the colocalization of the two markers. Nuclei was stained by <t>DAPI</t> (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (K, L). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (M, N). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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.)
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    SouthernBiotech microscope slides
    Ferritinophagy aggravated MV-induced pulmonary fibrosis through regulating ferroptosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ferritinophagy markers NCOA4 and FTH in MLE-12 cells with and without MS. n = 3 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. (G, H). Immunoblot and quantitative analysis showing the expression levels of SLC7A11, GPX4 and FTH in DFO-treated MLE-12 cells with and without MS. n = 3 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (I), or FTH (Green) and NCOA4 (Red) in the lung tissues (J). White arrows point to the colocalization of the two markers. Nuclei was stained by <t>DAPI</t> (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (K, L). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (M, N). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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.)
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    Image Search Results


    (A) Experimental paradigm for spontaneous withdrawal consisting of 7 days of twice daily escalating doses of morphine (or saline control), followed by a single final dose and 24 hours of abstinence before tissue collection. (B) Example images of anti-Fos immunostaining after spontaneous withdrawal paradigm, counterstained with DAPI. Dotted lines indicate boundaries of MHb and LHb. Arrow indicates Fos + cluster in the LHb. (C) Average distribution of Fos + cells across anterior-posterior axis of the habenula for each treatment condition. Contour lines indicate density of Fos + cells, normalized to the maximum density in the saline condition. (D) Quantification of the density of Fos + neurons in saline control and spontaneous withdrawal conditions. Each circle represents the mean for an individual animal. n=186 sections total from 10 mice per condition. **p< 0.01 by t-test. (E) Quantification of Fos + cells in all sections, binned every 400 μm along the A-P axis. Each circle represents one tissue section. (F) Same data in D, separated by sex. Two-Way ANOVA, treatment effect, **p< 0.01; sex effect, n.s.; interaction, n.s. Fisher’s post hoc, **p< 0.01. Open circles indicate female, filled circles indicate male.

    Journal: bioRxiv

    Article Title: Opioid withdrawal engages a habenular subpopulation responsive to aversive states

    doi: 10.64898/2026.04.29.721196

    Figure Lengend Snippet: (A) Experimental paradigm for spontaneous withdrawal consisting of 7 days of twice daily escalating doses of morphine (or saline control), followed by a single final dose and 24 hours of abstinence before tissue collection. (B) Example images of anti-Fos immunostaining after spontaneous withdrawal paradigm, counterstained with DAPI. Dotted lines indicate boundaries of MHb and LHb. Arrow indicates Fos + cluster in the LHb. (C) Average distribution of Fos + cells across anterior-posterior axis of the habenula for each treatment condition. Contour lines indicate density of Fos + cells, normalized to the maximum density in the saline condition. (D) Quantification of the density of Fos + neurons in saline control and spontaneous withdrawal conditions. Each circle represents the mean for an individual animal. n=186 sections total from 10 mice per condition. **p< 0.01 by t-test. (E) Quantification of Fos + cells in all sections, binned every 400 μm along the A-P axis. Each circle represents one tissue section. (F) Same data in D, separated by sex. Two-Way ANOVA, treatment effect, **p< 0.01; sex effect, n.s.; interaction, n.s. Fisher’s post hoc, **p< 0.01. Open circles indicate female, filled circles indicate male.

    Article Snippet: Sections were washed in 1xPBS and mounted on coverslips with Fluoromount-G with DAPI (Fisher Scientific, 50-112-8966).

    Techniques: Saline, Control, Immunostaining

    (A) Immgen ULI RNA sequencing data showing expression of Mrgpra6 in a subset of immune cells. (B) Bulk RNA sequencing of sorted lung basophils showing normalized transcripts per million (TPM) expression of highly expressed basophil genes Cd200R3, Mcpt8, and Il4 compared to Mrgpra6 (n = 5). (C) Flow cytometry gating of live, CD45 + blood cells identifying the relative contribution of basophils, eosinophils, neutrophils, and mast cell progenitors to the tdTomato + population. (D) Representative confocal images of sections of the lung from Mrgpra6CreER; Rosa26LSL-tdTomato mice showing DAPI (blue) and tdTomato (red) expression in Mrgpra6-expressing cells. (E-F) Flow cytometric analysis of lung basophils demonstrates that ∼80% are tdTomato + , consistent with Mrgpra6 expression in this population (E). Conversely, ∼80% of lung tdTomato+ cells are Cd200r3+ FcerI+ basophils, supporting the specificity of Mrgpra6 to basophils in the lung (F) (n = 9).

    Journal: bioRxiv

    Article Title: A basophil-specific GPCR mediates the immune response to helminth infection

    doi: 10.64898/2026.04.06.716327

    Figure Lengend Snippet: (A) Immgen ULI RNA sequencing data showing expression of Mrgpra6 in a subset of immune cells. (B) Bulk RNA sequencing of sorted lung basophils showing normalized transcripts per million (TPM) expression of highly expressed basophil genes Cd200R3, Mcpt8, and Il4 compared to Mrgpra6 (n = 5). (C) Flow cytometry gating of live, CD45 + blood cells identifying the relative contribution of basophils, eosinophils, neutrophils, and mast cell progenitors to the tdTomato + population. (D) Representative confocal images of sections of the lung from Mrgpra6CreER; Rosa26LSL-tdTomato mice showing DAPI (blue) and tdTomato (red) expression in Mrgpra6-expressing cells. (E-F) Flow cytometric analysis of lung basophils demonstrates that ∼80% are tdTomato + , consistent with Mrgpra6 expression in this population (E). Conversely, ∼80% of lung tdTomato+ cells are Cd200r3+ FcerI+ basophils, supporting the specificity of Mrgpra6 to basophils in the lung (F) (n = 9).

    Article Snippet: Sections were washed with PBST (0.3% Triton X-100), then mounted with Fluoromount-G with DAPI (ThermoFisher).

    Techniques: RNA Sequencing, Expressing, Flow Cytometry

    Ferritinophagy aggravated MV-induced pulmonary fibrosis through regulating ferroptosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ferritinophagy markers NCOA4 and FTH in MLE-12 cells with and without MS. n = 3 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. (G, H). Immunoblot and quantitative analysis showing the expression levels of SLC7A11, GPX4 and FTH in DFO-treated MLE-12 cells with and without MS. n = 3 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (I), or FTH (Green) and NCOA4 (Red) in the lung tissues (J). White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (K, L). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (M, N). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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.)

    Journal: Journal of Advanced Research

    Article Title: NCOA4-Mediated Ferritinophagy Induces Ferroptosis and Enriches Ferritin-Containing EVs via Ferritin Phase Separation to Promote Mechanical Ventilation-Induced Pulmonary Fibrosis

    doi: 10.1016/j.jare.2025.07.043

    Figure Lengend Snippet: Ferritinophagy aggravated MV-induced pulmonary fibrosis through regulating ferroptosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ferritinophagy markers NCOA4 and FTH in MLE-12 cells with and without MS. n = 3 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. (G, H). Immunoblot and quantitative analysis showing the expression levels of SLC7A11, GPX4 and FTH in DFO-treated MLE-12 cells with and without MS. n = 3 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (I), or FTH (Green) and NCOA4 (Red) in the lung tissues (J). White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (K, L). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (M, N). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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 slides were then washed and mounted using DAPI Fluoromount-G (0100–20, Southern Biotechnology, USA).

    Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Marker, Fluorescence

    Activation of ANG II/AGTR1 pathway initiated NCOA4-mediated ferritinophagy to aggravate MV-induced pulmonary fibrosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ANG II and AGTR1 in the lung homogenates. n = 4–6 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (G), or FTH (Green) and NCOA4 (Red) in the lung tissues (H). White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (I, J) Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (K, L). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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.)

    Journal: Journal of Advanced Research

    Article Title: NCOA4-Mediated Ferritinophagy Induces Ferroptosis and Enriches Ferritin-Containing EVs via Ferritin Phase Separation to Promote Mechanical Ventilation-Induced Pulmonary Fibrosis

    doi: 10.1016/j.jare.2025.07.043

    Figure Lengend Snippet: Activation of ANG II/AGTR1 pathway initiated NCOA4-mediated ferritinophagy to aggravate MV-induced pulmonary fibrosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of ANG II and AGTR1 in the lung homogenates. n = 4–6 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (E, F). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (G), or FTH (Green) and NCOA4 (Red) in the lung tissues (H). White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (I, J) Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (K, L). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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 slides were then washed and mounted using DAPI Fluoromount-G (0100–20, Southern Biotechnology, USA).

    Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining, Marker, Fluorescence

    Suppression of NCOA4 inhibited ferroptosis and alleviated MV-induced pulmonary fibrosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (E), or FTH (Green) and NCOA4 (Red) (F) in the lung tissues. White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (G, H). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (I, J). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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.)

    Journal: Journal of Advanced Research

    Article Title: NCOA4-Mediated Ferritinophagy Induces Ferroptosis and Enriches Ferritin-Containing EVs via Ferritin Phase Separation to Promote Mechanical Ventilation-Induced Pulmonary Fibrosis

    doi: 10.1016/j.jare.2025.07.043

    Figure Lengend Snippet: Suppression of NCOA4 inhibited ferroptosis and alleviated MV-induced pulmonary fibrosis. (A, B). Immunoblot and quantitative analysis showing the expression levels of NCOA4 and FTH in the lung homogenates. n = 4–6 per group. (C, D). Immunoblot and quantitative analysis showing the expression levels of SLC7A11 and GPX4 in the lung homogenates. n = 4–6 per group. Representative immunofluorescence staining images and relevant quantitative analysis for the epithelial marker E-cadherin (Green) and NCOA4 (Red) (E), or FTH (Green) and NCOA4 (Red) (F) in the lung tissues. White arrows point to the colocalization of the two markers. Nuclei was stained by DAPI (blue). Scale bars correspond to 20 μm. MFI, mean fluorescence intensity. (G, H). Immunoblot and quantitative analysis showing the expression levels of fibronectin and α-SMA in the lung homogenates. n = 4–6 per group. (I, J). Representative histopathologic images of H&E and Masson’s staining comparing the lung injury and collagen deposition. Original magnification x 200. Scale bars correspond to 100 μm. All data are presented as mean ± SEM. * 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 slides were then washed and mounted using DAPI Fluoromount-G (0100–20, Southern Biotechnology, USA).

    Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Marker, Fluorescence

    Mechanical ventilation enriched EVs to promote activation of lung fibroblast through iron overload. (A). GO enrichment analysis of differential expressed genes in the type II alveolar epithelial cells and fibroblasts originated from the MV group versus the control group. (B). Immunoblot analysis of the characteristic markers, Alix and CD63, and ferritin of EVs from control-BALF and MV-BALF. n = 3 per group. (C). NTA analysis of the concentration and size distribution of EVs from control-BALF and MV-BALF. (D). TEM images of EVs from control-BALF and MV-BALF. Original magnification x 80,000. Scale bars correspond to 200 nm. (E). Heatmap of the correlation strength of cellular interactions between different cells. (F). Cellchat analysis showing the intracellular interaction strength between AT2 or fibroblasts and other cell types in the MV group. (G, H). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with EVs. n = 3 per group. (I). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). Scale bars correspond to 10 μm, and n = 3 biologically independent samples. All data are presented as mean ± SEM. * 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.)

    Journal: Journal of Advanced Research

    Article Title: NCOA4-Mediated Ferritinophagy Induces Ferroptosis and Enriches Ferritin-Containing EVs via Ferritin Phase Separation to Promote Mechanical Ventilation-Induced Pulmonary Fibrosis

    doi: 10.1016/j.jare.2025.07.043

    Figure Lengend Snippet: Mechanical ventilation enriched EVs to promote activation of lung fibroblast through iron overload. (A). GO enrichment analysis of differential expressed genes in the type II alveolar epithelial cells and fibroblasts originated from the MV group versus the control group. (B). Immunoblot analysis of the characteristic markers, Alix and CD63, and ferritin of EVs from control-BALF and MV-BALF. n = 3 per group. (C). NTA analysis of the concentration and size distribution of EVs from control-BALF and MV-BALF. (D). TEM images of EVs from control-BALF and MV-BALF. Original magnification x 80,000. Scale bars correspond to 200 nm. (E). Heatmap of the correlation strength of cellular interactions between different cells. (F). Cellchat analysis showing the intracellular interaction strength between AT2 or fibroblasts and other cell types in the MV group. (G, H). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with EVs. n = 3 per group. (I). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). Scale bars correspond to 10 μm, and n = 3 biologically independent samples. All data are presented as mean ± SEM. * 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 slides were then washed and mounted using DAPI Fluoromount-G (0100–20, Southern Biotechnology, USA).

    Techniques: Activation Assay, Control, Western Blot, Concentration Assay, Confocal Microscopy, Staining

    MV induced ferritinophagy promoted the transfer of iron from EVs to activate lung fibroblast. (A, B). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with BALF-EVs generated from chloroquine intervened mice. n = 3 per group. (C). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). (D). Immunofluorescence of α-SMA (red) in MRC-5 cells treated with BALF-EVs generated from chloroquine intervened mice. Nuclei was stained by DAPI (blue). (E, F). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with BALF-EVs generated from NCOA4-knockdown mice administrated by AAV. n = 3 per group. (G). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). (H). Immunofluorescence of α-SMA (red) in MRC-5 cells treated with BALF-EVs generated from NCOA4-knockdown mice administrated by AAV. Nuclei was stained by DAPI (blue). Scale bars correspond to 10 μm, and n = 3 biologically independent samples. All data are presented as mean ± SEM. * 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.)

    Journal: Journal of Advanced Research

    Article Title: NCOA4-Mediated Ferritinophagy Induces Ferroptosis and Enriches Ferritin-Containing EVs via Ferritin Phase Separation to Promote Mechanical Ventilation-Induced Pulmonary Fibrosis

    doi: 10.1016/j.jare.2025.07.043

    Figure Lengend Snippet: MV induced ferritinophagy promoted the transfer of iron from EVs to activate lung fibroblast. (A, B). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with BALF-EVs generated from chloroquine intervened mice. n = 3 per group. (C). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). (D). Immunofluorescence of α-SMA (red) in MRC-5 cells treated with BALF-EVs generated from chloroquine intervened mice. Nuclei was stained by DAPI (blue). (E, F). Immunoblot and quantitative analysis of fibronectin, α-SMA and FTH in MRC-5 cells treated with BALF-EVs generated from NCOA4-knockdown mice administrated by AAV. n = 3 per group. (G). Representative images of ferrous iron (red) in the MRC-5 cells treated with PKH-67 labelled EVs (green) by the confocal microscopy. Nuclei was stained by DAPI (blue). (H). Immunofluorescence of α-SMA (red) in MRC-5 cells treated with BALF-EVs generated from NCOA4-knockdown mice administrated by AAV. Nuclei was stained by DAPI (blue). Scale bars correspond to 10 μm, and n = 3 biologically independent samples. All data are presented as mean ± SEM. * 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 slides were then washed and mounted using DAPI Fluoromount-G (0100–20, Southern Biotechnology, USA).

    Techniques: Western Blot, Generated, Confocal Microscopy, Staining, Immunofluorescence, Knockdown