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transit lt1 transfection reagent  (Mirus Bio)


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    Structured Review

    Mirus Bio transit lt1 transfection reagent
    Transit Lt1 Transfection Reagent, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 99/100, based on 8554 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transit+-mrna/TransIT+-LT1/pm42906373-172-63-66
    Average 99 stars, based on 8554 article reviews
    transit lt1 transfection reagent - by Bioz Stars, 2026-11
    99/100 stars

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    Related Articles

    Luciferase:

    Article Title: Pilot Proteomic Analysis of Urinary Extracellular Vesicles Supports the “Toxic Urine Hypothesis” as a Vicious Cycle in Refractory IC/BPS Pathogenesis
    Article Snippet: .. After 16 h, cells were co-transfected with pGL3-NF-κB firefly luciferase reporter (NovoPro Bioscience Inc., Shanghai, China) and pRL-TK Renilla control using TransIT (Mirus Bio LLC, Madison, WI, USA). ..

    Article Title: Pilot Proteomic Analysis of Urinary Extracellular Vesicles Supports the “Toxic Urine Hypothesis” as a Vicious Cycle in Refractory IC/BPS Pathogenesis
    Article Snippet: .. After 16 h, cells were co-transfected with pGL3-NF-κB firefly luciferase reporter (NovoPro Bioscience Inc., Shanghai, China) and pRL-TK Renilla control using TransIT (Mirus Bio LLC, Madison, WI, USA). ..

    Control:

    Article Title: Pilot Proteomic Analysis of Urinary Extracellular Vesicles Supports the “Toxic Urine Hypothesis” as a Vicious Cycle in Refractory IC/BPS Pathogenesis
    Article Snippet: .. After 16 h, cells were co-transfected with pGL3-NF-κB firefly luciferase reporter (NovoPro Bioscience Inc., Shanghai, China) and pRL-TK Renilla control using TransIT (Mirus Bio LLC, Madison, WI, USA). ..

    Article Title: Pilot Proteomic Analysis of Urinary Extracellular Vesicles Supports the “Toxic Urine Hypothesis” as a Vicious Cycle in Refractory IC/BPS Pathogenesis
    Article Snippet: .. After 16 h, cells were co-transfected with pGL3-NF-κB firefly luciferase reporter (NovoPro Bioscience Inc., Shanghai, China) and pRL-TK Renilla control using TransIT (Mirus Bio LLC, Madison, WI, USA). ..

    Transfection:

    Article Title: Evolution of a fuzzy ribonucleoprotein complex in viral assembly
    Article Snippet: Plasmids Cov2-N (1.34), CoV2-M-IRES-E (0.659), CoV-2- Spike (0.0032) and Luc-T20 (2.0) at indicated mass ratios for a total of 4 μg of DNA were diluted in 250 μL Opti-MEM reduced serum medium (Gibco cat# 31985062) at room temperature. .. 12 μL of TransIT–293 Transfection Reagent (Mirus Bio, cat# MIR 2704) equilibrated to room temperature was added to each sample, and immediately vortexed. ..

    Article Title: Cell-cell fusion limits activation of the unfolded protein response induced by the Nipah virus glycoproteins
    Article Snippet: After virus adsorption, cells were washed five times with DMEM 2% FCS and incubated in DMEM 2% FCS containing 500 nM thapsigargin (Tg; Sigma), 20 mM NH 4 Cl (Merck), or DMSO (Wak-Chemie) for 18–24 h. To determine infectious viral titers, the cell-free supernatants of infected cells were collected and quantified by serial dilution on Vero76 cells. .. Cells at 80% confluency were transfected using Lipofectamine 2000 (Invitrogen) or TransIT (Mirus Bio) according to the manufacturer’s protocol. ..

    Article Title: NicheSphere reveals Spp1⁺ macrophages as central hubs coordinating fibrotic remodeling in myeloproliferative neoplasms
    Article Snippet: .. Lentiviral particles were produced by transient transfection with lentiviral plasmid together with pSPAX and VSVG packaging plasmids using TransIT (Mirus). ..

    Article Title: The E3 ligases Itch and WWP2 regulate autoimmune neuroinflammation by controlling T H 2 to T H 17 cell conversion via interleukin-4-STAT5 axis in mice
    Article Snippet: .. The plasmid and pCL-ECO packaging vector were transfected into PLAT-E (Platinum-Eco)cells using TransIT (Mirus, #MIR 2154) according to the manufacturer’s instructions . ..

    Article Title: An mRNA vaccine encoding the Ebola virus glycoprotein induces high neutralizing antibody titers and provides strong protection against lethal infections in mouse models
    Article Snippet: .. A549 cells were seeded, rested for 24 h, and transfected with 2 μg of mRNA using Lipofectamine MessengerMAX (Invitrogen, Waltham, MA) or TransIT (Mirus Bio, Madison, WI) transfection reagent according to the manufacturers’ direction. ..

    Article Title: The core herpes simplex-1 fusion complex drives cell-to-cell spreading of pathological Tau
    Article Snippet: The Vi-CELL XR Cell Viability Analyzer (Beckman Coulter) was used to determine the total numbers of viable cells. .. Cells were transfected using TransIT-2020/X2 (Mirus) or Lipofectamine 2000 (Invitrogen) reagents according to the manufacturer’s instructions. .. Competent E. coli BL21 (DE3) cells were transformed with a pET vector coding for the Saccharomyces cerevisiae Sup35 NM domain with a C-terminal His-tag under the control of the T7 promoter, at a concentration of 100 ng.

    Produced:

    Article Title: NicheSphere reveals Spp1⁺ macrophages as central hubs coordinating fibrotic remodeling in myeloproliferative neoplasms
    Article Snippet: .. Lentiviral particles were produced by transient transfection with lentiviral plasmid together with pSPAX and VSVG packaging plasmids using TransIT (Mirus). ..

    Plasmid Preparation:

    Article Title: NicheSphere reveals Spp1⁺ macrophages as central hubs coordinating fibrotic remodeling in myeloproliferative neoplasms
    Article Snippet: .. Lentiviral particles were produced by transient transfection with lentiviral plasmid together with pSPAX and VSVG packaging plasmids using TransIT (Mirus). ..

    Article Title: The E3 ligases Itch and WWP2 regulate autoimmune neuroinflammation by controlling T H 2 to T H 17 cell conversion via interleukin-4-STAT5 axis in mice
    Article Snippet: .. The plasmid and pCL-ECO packaging vector were transfected into PLAT-E (Platinum-Eco)cells using TransIT (Mirus, #MIR 2154) according to the manufacturer’s instructions . ..



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    Genetic and Pharmacological Inhibition of Nupr1 Attenuates Ang II Induced Mitochondrial Apoptosis in VSMCs. (A) Cell ability was evaluated by CCK8 assay ( n = 8 per group). (B) Lactate production of indicated groups ( n = 6 per group). (C) Representative images of TEM in VSMCs from the indicated groups. (D) Quantification of mitochondrial size in VSMCs from indicated groups ( n = 6 per group). (E) Quantification of circularity index in VSMC from indicated groups ( n = 6 per group). (F) Real‐time monitoring of oxygen consumption rate (OCR) in indicated groups detected by O2k. (G–I) Quantification of ATP‐coupled respiration, mitochondrial basal respiration, and maximal respiration from indicated groups ( n = 6 per group). (J) Representative images of <t>calcein</t> and Annexin V fluorescence results showing <t>mPTP</t> opening and apoptosis in indicated groups. Stronger green fluorescence represents a lower degree of mPTP opening, while diminished fluorescence indicates increased mPTP opening. Stronger red fluorescence represents a higher apoptosis rate. (K, L) Quantification of fluorescence intensity, including Annexin V and Calcein ( n = 6 per group). (M) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (N) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group). (O) Quantification of maximal respiration from indicated groups ( n = 6 per group). (P) Real‐time monitoring of OCR in indicated groups detected by O2k. (Q, R) Quantification of ATP‐coupled respiration and mitochondrial basal respiration from indicated groups ( n = 6 per group). (S) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (T) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group).
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    Beyotime mptp assay kit
    Increased mechanical stiffness induces <t>MPTP</t> opening and mitochondrial damage in macrophages. (A) MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial membrane potential was labeled using the mitochondrial membrane potential probe JC‐1 and detected by flow cytometry, n = 6 biological replicates. (B). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial reactive oxygen species content in cells was measured by using the mitoSOX probe and detected by flow cytometry, n = 5 biological replicates. (C). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Calcium ion concentration in cytoplasm was detected using flow cytometry, n = 5 biological replicates. (D). MH‐S cells were incubated with different stiffness matrix gels for 24 h. MPTP activity of macrophages after matrix gel culture was assayed using the MPTP kit, scale bar = 100 µm, n = 6 biological replicates, stronger green fluorescence corresponds to reduced MPTP opening and preserved mitochondrial integrity, while weaker fluorescence indicates elevated MPTP opening and impaired mitochondrial structural integrity. (E). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of mitochondrial membrane permeability transition pore‐associated genes ANT1 and CypD detected by qPCR, n = 5 biological replicates. (F). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of ANT1 and CypD was detected by WB, n = 5 biological replicates. (G). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of CD163 and iNOS was detected by WB, n = 6 biological replicates.
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    Image Search Results


    Genetic and Pharmacological Inhibition of Nupr1 Attenuates Ang II Induced Mitochondrial Apoptosis in VSMCs. (A) Cell ability was evaluated by CCK8 assay ( n = 8 per group). (B) Lactate production of indicated groups ( n = 6 per group). (C) Representative images of TEM in VSMCs from the indicated groups. (D) Quantification of mitochondrial size in VSMCs from indicated groups ( n = 6 per group). (E) Quantification of circularity index in VSMC from indicated groups ( n = 6 per group). (F) Real‐time monitoring of oxygen consumption rate (OCR) in indicated groups detected by O2k. (G–I) Quantification of ATP‐coupled respiration, mitochondrial basal respiration, and maximal respiration from indicated groups ( n = 6 per group). (J) Representative images of calcein and Annexin V fluorescence results showing mPTP opening and apoptosis in indicated groups. Stronger green fluorescence represents a lower degree of mPTP opening, while diminished fluorescence indicates increased mPTP opening. Stronger red fluorescence represents a higher apoptosis rate. (K, L) Quantification of fluorescence intensity, including Annexin V and Calcein ( n = 6 per group). (M) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (N) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group). (O) Quantification of maximal respiration from indicated groups ( n = 6 per group). (P) Real‐time monitoring of OCR in indicated groups detected by O2k. (Q, R) Quantification of ATP‐coupled respiration and mitochondrial basal respiration from indicated groups ( n = 6 per group). (S) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (T) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group).

    Journal: Advanced Science

    Article Title: Targeting Nupr1‐Mediated p53 Mitochondrial Sequestration Alleviates Aortic Dissection by Restoring the Gadd45b‐Dependent Survival Program

    doi: 10.1002/advs.78143

    Figure Lengend Snippet: Genetic and Pharmacological Inhibition of Nupr1 Attenuates Ang II Induced Mitochondrial Apoptosis in VSMCs. (A) Cell ability was evaluated by CCK8 assay ( n = 8 per group). (B) Lactate production of indicated groups ( n = 6 per group). (C) Representative images of TEM in VSMCs from the indicated groups. (D) Quantification of mitochondrial size in VSMCs from indicated groups ( n = 6 per group). (E) Quantification of circularity index in VSMC from indicated groups ( n = 6 per group). (F) Real‐time monitoring of oxygen consumption rate (OCR) in indicated groups detected by O2k. (G–I) Quantification of ATP‐coupled respiration, mitochondrial basal respiration, and maximal respiration from indicated groups ( n = 6 per group). (J) Representative images of calcein and Annexin V fluorescence results showing mPTP opening and apoptosis in indicated groups. Stronger green fluorescence represents a lower degree of mPTP opening, while diminished fluorescence indicates increased mPTP opening. Stronger red fluorescence represents a higher apoptosis rate. (K, L) Quantification of fluorescence intensity, including Annexin V and Calcein ( n = 6 per group). (M) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (N) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group). (O) Quantification of maximal respiration from indicated groups ( n = 6 per group). (P) Real‐time monitoring of OCR in indicated groups detected by O2k. (Q, R) Quantification of ATP‐coupled respiration and mitochondrial basal respiration from indicated groups ( n = 6 per group). (S) Flow cytometry analysis of Annexin and PI staining of apoptosis from indicated groups. (T) Quantification of early apoptosis, late apoptosis, and total apoptosis by flow cytometry ( n = 3 per group).

    Article Snippet: According to mPTP test kit instructions (C2009S, Beyotime), 200 μL of Calcein AM staining solution and fluorescence quenching working solution were added to each well after the respective cell treatments.

    Techniques: Inhibition, CCK-8 Assay, Real-time Monitoring, Fluorescence, Flow Cytometry, Analysis, Staining

    Gadd45b is Indispensable for the Protective Effects of Nupr1 Deficiency Against Mitochondrial Apoptosis. (A) Enrichment of KEGG pathways. (B) GSEA analysis in VSMCs treated with Ang II stimulation and Nupr1 deficiency. (C) Chordal diagram of GO pathways related with cell fate and the contributing DEPs. (D) Stem plot of DEPs in the apoptosis pathway from the indicated groups. (E) RT‐qPCR analysis of mRNA levels of candidate genes in VSMCs. Data were presented as relative fold change to controls ( n = 6 per group). (F) Expressions of Gadd45b in VSMCs measured by Western blot and its quantification from indicated groups measured by Western blot ( n = 5 per group). (G, H) Representative images of ROS staining in indicated groups. Semi‐quantitative analysis of mean fluorescence intensity in indicated groups ( n = 6 per group). (I) Representative images of calcein and Annexin V fluorescence results showing mPTP opening in indicated groups. (J) Quantification of fluorescence intensity, including Annexin V and mPTP ( n = 3 per group). (K) Co‐immunoprecipitation and Western blot assays of Myc‐Nupr1 and Gadd45b. (L) Co‐immunoprecipitation assays of Myc‐Gadd45b and Nupr1.

    Journal: Advanced Science

    Article Title: Targeting Nupr1‐Mediated p53 Mitochondrial Sequestration Alleviates Aortic Dissection by Restoring the Gadd45b‐Dependent Survival Program

    doi: 10.1002/advs.78143

    Figure Lengend Snippet: Gadd45b is Indispensable for the Protective Effects of Nupr1 Deficiency Against Mitochondrial Apoptosis. (A) Enrichment of KEGG pathways. (B) GSEA analysis in VSMCs treated with Ang II stimulation and Nupr1 deficiency. (C) Chordal diagram of GO pathways related with cell fate and the contributing DEPs. (D) Stem plot of DEPs in the apoptosis pathway from the indicated groups. (E) RT‐qPCR analysis of mRNA levels of candidate genes in VSMCs. Data were presented as relative fold change to controls ( n = 6 per group). (F) Expressions of Gadd45b in VSMCs measured by Western blot and its quantification from indicated groups measured by Western blot ( n = 5 per group). (G, H) Representative images of ROS staining in indicated groups. Semi‐quantitative analysis of mean fluorescence intensity in indicated groups ( n = 6 per group). (I) Representative images of calcein and Annexin V fluorescence results showing mPTP opening in indicated groups. (J) Quantification of fluorescence intensity, including Annexin V and mPTP ( n = 3 per group). (K) Co‐immunoprecipitation and Western blot assays of Myc‐Nupr1 and Gadd45b. (L) Co‐immunoprecipitation assays of Myc‐Gadd45b and Nupr1.

    Article Snippet: According to mPTP test kit instructions (C2009S, Beyotime), 200 μL of Calcein AM staining solution and fluorescence quenching working solution were added to each well after the respective cell treatments.

    Techniques: Analysis, Quantitative RT-PCR, Western Blot, Staining, Fluorescence, Co-Immunoprecipitation Assay, Significance Assay

    Increased mechanical stiffness induces MPTP opening and mitochondrial damage in macrophages. (A) MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial membrane potential was labeled using the mitochondrial membrane potential probe JC‐1 and detected by flow cytometry, n = 6 biological replicates. (B). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial reactive oxygen species content in cells was measured by using the mitoSOX probe and detected by flow cytometry, n = 5 biological replicates. (C). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Calcium ion concentration in cytoplasm was detected using flow cytometry, n = 5 biological replicates. (D). MH‐S cells were incubated with different stiffness matrix gels for 24 h. MPTP activity of macrophages after matrix gel culture was assayed using the MPTP kit, scale bar = 100 µm, n = 6 biological replicates, stronger green fluorescence corresponds to reduced MPTP opening and preserved mitochondrial integrity, while weaker fluorescence indicates elevated MPTP opening and impaired mitochondrial structural integrity. (E). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of mitochondrial membrane permeability transition pore‐associated genes ANT1 and CypD detected by qPCR, n = 5 biological replicates. (F). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of ANT1 and CypD was detected by WB, n = 5 biological replicates. (G). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of CD163 and iNOS was detected by WB, n = 6 biological replicates.

    Journal: MedComm

    Article Title: The PIEZO1/CISD1/IP3R Mechanotransduction Axis Drives Macrophage Mitochondrial Dysfunction and Inflammatory Activation in Pulmonary Fibrosis

    doi: 10.1002/mco2.71080

    Figure Lengend Snippet: Increased mechanical stiffness induces MPTP opening and mitochondrial damage in macrophages. (A) MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial membrane potential was labeled using the mitochondrial membrane potential probe JC‐1 and detected by flow cytometry, n = 6 biological replicates. (B). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Mitochondrial reactive oxygen species content in cells was measured by using the mitoSOX probe and detected by flow cytometry, n = 5 biological replicates. (C). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Calcium ion concentration in cytoplasm was detected using flow cytometry, n = 5 biological replicates. (D). MH‐S cells were incubated with different stiffness matrix gels for 24 h. MPTP activity of macrophages after matrix gel culture was assayed using the MPTP kit, scale bar = 100 µm, n = 6 biological replicates, stronger green fluorescence corresponds to reduced MPTP opening and preserved mitochondrial integrity, while weaker fluorescence indicates elevated MPTP opening and impaired mitochondrial structural integrity. (E). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of mitochondrial membrane permeability transition pore‐associated genes ANT1 and CypD detected by qPCR, n = 5 biological replicates. (F). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of ANT1 and CypD was detected by WB, n = 5 biological replicates. (G). MH‐S cells were incubated with different stiffness matrix gels for 24 h. Expression of CD163 and iNOS was detected by WB, n = 6 biological replicates.

    Article Snippet: MPTP activity was detected using the MPTP assay kit (Beyotime Biotech, Shanghai, China).

    Techniques: Incubation, Membrane, Labeling, Flow Cytometry, Concentration Assay, Activity Assay, Fluorescence, Expressing, Permeability

    Knockdown of CISD1 attenuates mechanical stiffness‐mediated macrophage MPTP opening and mitochondrial damage. (A). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and cytosolic calcium ions were detected by flow cytometry, n = 5 biological replicates. (B). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and MPTP activity was detected by flow cytometry, n = 5 biological replicates. (C). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 5 biological replicates. (D and E) MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, mitochondrial membrane potential was detected by immunofluorescence microscope, n = 3 biological replicates, scale bar = 20 µm. (F and G). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and MPTP‐related proteins and inflammation‐related proteins were detected by WB, n = 4 biological replicates.

    Journal: MedComm

    Article Title: The PIEZO1/CISD1/IP3R Mechanotransduction Axis Drives Macrophage Mitochondrial Dysfunction and Inflammatory Activation in Pulmonary Fibrosis

    doi: 10.1002/mco2.71080

    Figure Lengend Snippet: Knockdown of CISD1 attenuates mechanical stiffness‐mediated macrophage MPTP opening and mitochondrial damage. (A). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and cytosolic calcium ions were detected by flow cytometry, n = 5 biological replicates. (B). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and MPTP activity was detected by flow cytometry, n = 5 biological replicates. (C). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 5 biological replicates. (D and E) MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, mitochondrial membrane potential was detected by immunofluorescence microscope, n = 3 biological replicates, scale bar = 20 µm. (F and G). MH‐S cells with knockdown of CISD1 were cultured on different stiffness matrix gels, and MPTP‐related proteins and inflammation‐related proteins were detected by WB, n = 4 biological replicates.

    Article Snippet: MPTP activity was detected using the MPTP assay kit (Beyotime Biotech, Shanghai, China).

    Techniques: Knockdown, Cell Culture, Flow Cytometry, Activity Assay, Membrane, Immunofluorescence, Microscopy

    Matrix stiffness upregulates CISD1 via PIEZO1 to mediate mitochondrial damage. (A) MH‐S cells were treated with 2.5 µM or 5 µM Yoda1 for 24 h. WB assay was performed to detect the expression of MPTP‐related proteins, n = 6 biological replicates. (B) MH‐S cells were treated with 5 µM Yoda1 for 24 h, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 6 biological replicates. (C) MH‐S cells were treated with 5 µM Yoda1 for 24 h, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 6 biological replicates, scale bar = 100 µm. (D) MH‐S cells were treated with 5 µM Yoda1 for 24 h, MPTP opening was detected by immunofluorescence microscopy, n = 6 biological replicates, scale bar = 100 µm. (E) MH‐S cells were cultured with matrix gels of different stiffness and simultaneously treated with PIEZO1 inhibitor GsMTx4, WB assay was performed to detect the expression of MPTP‐related proteins, n = 4 biological replicates. (F) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, and MPTP activity was detected by flow cytometry, n = 3 biological replicates. (G) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 5 biological replicates. (H) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 4 biological replicates, scale bar = 20 µm.

    Journal: MedComm

    Article Title: The PIEZO1/CISD1/IP3R Mechanotransduction Axis Drives Macrophage Mitochondrial Dysfunction and Inflammatory Activation in Pulmonary Fibrosis

    doi: 10.1002/mco2.71080

    Figure Lengend Snippet: Matrix stiffness upregulates CISD1 via PIEZO1 to mediate mitochondrial damage. (A) MH‐S cells were treated with 2.5 µM or 5 µM Yoda1 for 24 h. WB assay was performed to detect the expression of MPTP‐related proteins, n = 6 biological replicates. (B) MH‐S cells were treated with 5 µM Yoda1 for 24 h, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 6 biological replicates. (C) MH‐S cells were treated with 5 µM Yoda1 for 24 h, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 6 biological replicates, scale bar = 100 µm. (D) MH‐S cells were treated with 5 µM Yoda1 for 24 h, MPTP opening was detected by immunofluorescence microscopy, n = 6 biological replicates, scale bar = 100 µm. (E) MH‐S cells were cultured with matrix gels of different stiffness and simultaneously treated with PIEZO1 inhibitor GsMTx4, WB assay was performed to detect the expression of MPTP‐related proteins, n = 4 biological replicates. (F) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, and MPTP activity was detected by flow cytometry, n = 3 biological replicates. (G) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 5 biological replicates. (H) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with PIEZO1 inhibitor GsMTx4, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 4 biological replicates, scale bar = 20 µm.

    Article Snippet: MPTP activity was detected using the MPTP assay kit (Beyotime Biotech, Shanghai, China).

    Techniques: Expressing, Flow Cytometry, Membrane, Immunofluorescence, Microscopy, Cell Culture, Activity Assay

    Matrix stiffness‐mediated mitochondrial damage depends on IP3R activity. (A) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, and cytosolic calcium ions were detected by flow cytometry, n = 3 biological replicates. (B) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, and MPTP activity was detected by flow cytometry, n = 3 biological replicates. (C) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 3 biological replicates. (D) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 3 biological replicates, scale bar = 20 µm.

    Journal: MedComm

    Article Title: The PIEZO1/CISD1/IP3R Mechanotransduction Axis Drives Macrophage Mitochondrial Dysfunction and Inflammatory Activation in Pulmonary Fibrosis

    doi: 10.1002/mco2.71080

    Figure Lengend Snippet: Matrix stiffness‐mediated mitochondrial damage depends on IP3R activity. (A) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, and cytosolic calcium ions were detected by flow cytometry, n = 3 biological replicates. (B) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, and MPTP activity was detected by flow cytometry, n = 3 biological replicates. (C) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, mitochondrial reactive oxygen species content was detected by flow cytometry, n = 3 biological replicates. (D) MH‐S cells were cultured with different stiffness matrix gels and simultaneously treated with IP3R inhibitor 2‐APB, mitochondrial membrane potential was detected by immunofluorescence microscopy, n = 3 biological replicates, scale bar = 20 µm.

    Article Snippet: MPTP activity was detected using the MPTP assay kit (Beyotime Biotech, Shanghai, China).

    Techniques: Activity Assay, Cell Culture, Flow Cytometry, Membrane, Immunofluorescence, Microscopy