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anti mff  (Proteintech)


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    Proteintech anti mff
    Anti Mff, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+mff/MFF+Antibody/10__1038_slash_s41467___026___78424___w-288-61-62
    Average 96 stars, based on 245 article reviews
    anti mff - by Bioz Stars, 2026-11
    96/100 stars

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    Related Articles

    other:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation
    Article Snippet: Proteins in cell lysate were quantified using Pierce BCA Protein Assay Kit.

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation
    Article Snippet: Total proteins were extracted from human trophoblast HTR-8/SVneo cells, human villous tissues, and mouse placental tissues using RIPA lysis buffer (Thermo Fisher Scientific) and quantified using Pierce BCA Protein Assay Kit (Pierce).

    Control:

    Article Title: Mitorubin, berberrubine-based compounds that improve mitochondrial function, exhibit cardioprotective effects against age-related cardiac dysfunction.
    Article Snippet: C2C12 cells were transfected using Lipofectamine 3000 (Thermo Fisher Scientific), and knockout clones were selected by serial dilution. .. The following antibodies were used: anti-MITOL (LS-C164034, LSBio); anti-α-Tubulin (T-9026, Sigma-Aldrich); anti-Mfn2 (sc-100560, Santa Cruz Biotechnology); anti-HSP60 (sc-136291, Santa Cruz Biotechnology); anti-ATP5a (ab14748, Abcam); anti-OPA1 (612606, BD Biosciences); anti-Fis1 (GTX111010, GeneTex); anti-MFF (17090-1-AP, Proteintech); anti-4HNE (ab46545, Abcam) for cellbased assays; anti-4HNE (clone HNEJ-2, MHN, Japan Institute for the Control of Aging (JaICA), NIKKEN SEIL) for tissue samples; anti-LC3A/B (D3U4C) (12741, Cell Signaling Technology); anti-SQSTM1/p62 (D6M5X) (23214, Cell Signaling Technology); anti-Tom20 (11802-1-AP, Proteintech); and anti-Vinculin (V9131, Sigma-Aldrich). .. Proteins in sample buffer were separated by SDS-PAGE and transferred to Immobilon-P PVDF membranes (IPVH00010, Millipore).

    Lysis:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: .. Cells and tissues lysates (1 × 10 7 cells in 0.1 mL IP lysis buffer or 0.2 g tissues in 1 mL IP lysis buffer) was incubated with anti-MFF (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463) at 4 ◦ C overnight with gentle rotation, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931) as negative control, and then incubated with 40 μL Protein A/G Magnetic Beads (B23201, MCE, USA) at 4 ◦ C for 4 h with gentle rotation. ..

    Incubation:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: .. Cells and tissues lysates (1 × 10 7 cells in 0.1 mL IP lysis buffer or 0.2 g tissues in 1 mL IP lysis buffer) was incubated with anti-MFF (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463) at 4 ◦ C overnight with gentle rotation, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931) as negative control, and then incubated with 40 μL Protein A/G Magnetic Beads (B23201, MCE, USA) at 4 ◦ C for 4 h with gentle rotation. ..

    Article Title: CUMS stress facilitates hippocampal neural mitophagy through FIS1/MFF-mediated mitochondrial fragmentation.
    Article Snippet: Each sample was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membrane (Millipore). .. After being blocked in BSA/Tris-buffered saline with Tween 20 (TBST) at room temperature for 2 h, the membranes were incubated with anti-LC3 (1:1,000, 14600-1-AP, Proteintech, PR China), anti-Parkin (1:1,000, 1406 0-1-AP, Proteintech, PR China), anti-PINK1 (1:1,000, 23274-1-AP, Proteintech, PR China), antiPGC1a (1:5,000, 66369-Ig, Proteintech, PR China), anti-FIS1 (1:2,000, 66635-1-Ig, Proteintech, PR China), anti-MFF (1:2,000, 66527-1-Ig, Proteintech, PR China), anti-DRP1 (1:2,000, 12957-1- AP, Proteintech, PR China), and anti-GAPDH [1:8,000, horseradish peroxidase (HRP)-60004, Proteintech, PR China] at 4 C overnight, after the incubation of secondary antibodies, images were obtained by using enhanced chemiluminescence (ECL) and results were scanned by using the Gel imaging System (UVP Company, Upland, CA). ..

    Article Title: CUMS stress facilitates hippocampal neural mitophagy through FIS1/MFF-mediated mitochondrial fragmentation.
    Article Snippet: .. Slides were incubated overnight at 4 C in primary antibodies: anti-FIS1 (1:200, 66635-1-Ig, Proteintech, PR China), anti-MFF (1:200, 66527-1- Ig, Proteintech, PR China) and anti-DRP1 (1:250, 12957-1-AP, Proteintech, PR China) antibodies diluted in blocking buffer. .. Following three washes in PBS, sections were then incubated with secondary antibodies, Goat Anti-Mouse IgG (H&L)AbFluor 488 (Invitrogen, RS3208, 1:500) and Goat AntiRabbit IgG (H&L)-AbFluor 647 (Invitrogen, RS3811, 1:500) for 2h at room temperature.

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: Proteins (10–30 μg/well, equal amounts within group but different amounts among groups for better comparison) were separated on 10% or 12% SDS-PAGE gel and transferred on an equilibrated polyvinylidene difluoride membrane (PVDF, Amersham Biosciences, Buckinghamshire, 8 www.thelancet.com Vol 125 March, 2026 UK). .. After blocking with 5% bovine serum albumin (BSA, Sigma–Aldrich) in TBST (10 mM Tris–HCl, 150 mM NaCl, and 0.1% Tween 20) for 1 h at room temperature, the membrane was incubated with primary antibody overnight at 4 ◦ C. The primary antibodies contained anti-MFF (17090-1-AP, proteintech, dilution 1:1000, RRID: AB_2142463), anti-TRIM21 (67136-1-Ig, proteintech, dilution 1:1000, RRID: AB_ 2882435), anti-GLB1 (66586-1-Ig, proteintech, dilution 1:1000, RRID: AB_2881946), anti-p38 (80821-2-RR, proteintech, dilution 1:1000, RRID: AB_3670489), antip21 (ab109199, abcam, dilution 1:1000, RRID: AB_ 10861551), anti-p16 (ab189034, abcam, dilution 1:1000, RRID: RRID: AB_2737282), and anti-GAPDH (#5174, Cell Signalling Technology, dilution 1:1000, RRID: AB_10622025). ..

    Article Title: ACAT1/SOAT1 governs mitochondrial dynamics and thermogenesis via the mTOR-MFN2-USP2 axis in brown adipocytes
    Article Snippet: Proteins were separated by SDS-PAGE and transferred electrophoretically to a PVDF membrane (Sigma-Aldrich, 24937-79-9). .. Membranes were incubated with primary antibodies overnight at 4 °C: anti-ACAT1 (Purified by our lab), anti-HSP90 (ProteinTech, 60318-1-Ig), anti-Beta Actin (ProteinTech, 66009-1-Ig), antiOXPHOS (Abcam, ab110413), anti-UCP1 (Abcam, ab10983), antiCalnexin (ProteinTech, 10427-2-AP), anti-IP3R (ProteinTech, 19962-1- AP), anti-FACL4 (Abcam, ab155282), anti-VDAC (ProteinTech, 10866-1- AP), anti-Tubulin (ProteinTech, 66031-1-Ig), anti-MFN2 (ProteinTech, 12186-1-AP), anti-MFN1 (ProteinTech, 66776-1-Ig), anti-OPA1 (ProteinTech, 27733-1-AP), anti-DRP1 (ProteinTech, 12957-1-AP), antipSer616-DRP1 (Cell Signaling, 3455), anti-MFF (ProteinTech, 17090-1- AP), anti-Flag (Sigma, F1804), anti-P4D1 (Santa Cruz, sc-8017), anti-HA (ProteinTech, 51064-1-AP), anti-mTOR (ProteinTech, 66888-1-Ig), antipSer2448-mTOR (Cell signaling, 5536), anti-pThr389-p70S6Kinase (Cell signaling, 9234), anti-USP2 (ProteinTech, 10392-1-AP) and antiGNPAT (ProteinTech, 14931-1-AP). .. After incubation with primary antibodies, membranes were washed with TBST and incubated with secondary antibodies conjugated to horseradish peroxidase (HRP; Epizyme, LF101 and LF102) at room temperature for 1 hour.

    Gentle:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: .. Cells and tissues lysates (1 × 10 7 cells in 0.1 mL IP lysis buffer or 0.2 g tissues in 1 mL IP lysis buffer) was incubated with anti-MFF (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463) at 4 ◦ C overnight with gentle rotation, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931) as negative control, and then incubated with 40 μL Protein A/G Magnetic Beads (B23201, MCE, USA) at 4 ◦ C for 4 h with gentle rotation. ..

    Negative Control:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: .. Cells and tissues lysates (1 × 10 7 cells in 0.1 mL IP lysis buffer or 0.2 g tissues in 1 mL IP lysis buffer) was incubated with anti-MFF (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463) at 4 ◦ C overnight with gentle rotation, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931) as negative control, and then incubated with 40 μL Protein A/G Magnetic Beads (B23201, MCE, USA) at 4 ◦ C for 4 h with gentle rotation. ..

    Magnetic Beads:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: .. Cells and tissues lysates (1 × 10 7 cells in 0.1 mL IP lysis buffer or 0.2 g tissues in 1 mL IP lysis buffer) was incubated with anti-MFF (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463) at 4 ◦ C overnight with gentle rotation, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931) as negative control, and then incubated with 40 μL Protein A/G Magnetic Beads (B23201, MCE, USA) at 4 ◦ C for 4 h with gentle rotation. ..

    Saline:

    Article Title: CUMS stress facilitates hippocampal neural mitophagy through FIS1/MFF-mediated mitochondrial fragmentation.
    Article Snippet: Each sample was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membrane (Millipore). .. After being blocked in BSA/Tris-buffered saline with Tween 20 (TBST) at room temperature for 2 h, the membranes were incubated with anti-LC3 (1:1,000, 14600-1-AP, Proteintech, PR China), anti-Parkin (1:1,000, 1406 0-1-AP, Proteintech, PR China), anti-PINK1 (1:1,000, 23274-1-AP, Proteintech, PR China), antiPGC1a (1:5,000, 66369-Ig, Proteintech, PR China), anti-FIS1 (1:2,000, 66635-1-Ig, Proteintech, PR China), anti-MFF (1:2,000, 66527-1-Ig, Proteintech, PR China), anti-DRP1 (1:2,000, 12957-1- AP, Proteintech, PR China), and anti-GAPDH [1:8,000, horseradish peroxidase (HRP)-60004, Proteintech, PR China] at 4 C overnight, after the incubation of secondary antibodies, images were obtained by using enhanced chemiluminescence (ECL) and results were scanned by using the Gel imaging System (UVP Company, Upland, CA). ..

    Imaging:

    Article Title: CUMS stress facilitates hippocampal neural mitophagy through FIS1/MFF-mediated mitochondrial fragmentation.
    Article Snippet: Each sample was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membrane (Millipore). .. After being blocked in BSA/Tris-buffered saline with Tween 20 (TBST) at room temperature for 2 h, the membranes were incubated with anti-LC3 (1:1,000, 14600-1-AP, Proteintech, PR China), anti-Parkin (1:1,000, 1406 0-1-AP, Proteintech, PR China), anti-PINK1 (1:1,000, 23274-1-AP, Proteintech, PR China), antiPGC1a (1:5,000, 66369-Ig, Proteintech, PR China), anti-FIS1 (1:2,000, 66635-1-Ig, Proteintech, PR China), anti-MFF (1:2,000, 66527-1-Ig, Proteintech, PR China), anti-DRP1 (1:2,000, 12957-1- AP, Proteintech, PR China), and anti-GAPDH [1:8,000, horseradish peroxidase (HRP)-60004, Proteintech, PR China] at 4 C overnight, after the incubation of secondary antibodies, images were obtained by using enhanced chemiluminescence (ECL) and results were scanned by using the Gel imaging System (UVP Company, Upland, CA). ..

    Blocking Assay:

    Article Title: CUMS stress facilitates hippocampal neural mitophagy through FIS1/MFF-mediated mitochondrial fragmentation.
    Article Snippet: .. Slides were incubated overnight at 4 C in primary antibodies: anti-FIS1 (1:200, 66635-1-Ig, Proteintech, PR China), anti-MFF (1:200, 66527-1- Ig, Proteintech, PR China) and anti-DRP1 (1:250, 12957-1-AP, Proteintech, PR China) antibodies diluted in blocking buffer. .. Following three washes in PBS, sections were then incubated with secondary antibodies, Goat Anti-Mouse IgG (H&L)AbFluor 488 (Invitrogen, RS3208, 1:500) and Goat AntiRabbit IgG (H&L)-AbFluor 647 (Invitrogen, RS3811, 1:500) for 2h at room temperature.

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: Proteins (10–30 μg/well, equal amounts within group but different amounts among groups for better comparison) were separated on 10% or 12% SDS-PAGE gel and transferred on an equilibrated polyvinylidene difluoride membrane (PVDF, Amersham Biosciences, Buckinghamshire, 8 www.thelancet.com Vol 125 March, 2026 UK). .. After blocking with 5% bovine serum albumin (BSA, Sigma–Aldrich) in TBST (10 mM Tris–HCl, 150 mM NaCl, and 0.1% Tween 20) for 1 h at room temperature, the membrane was incubated with primary antibody overnight at 4 ◦ C. The primary antibodies contained anti-MFF (17090-1-AP, proteintech, dilution 1:1000, RRID: AB_2142463), anti-TRIM21 (67136-1-Ig, proteintech, dilution 1:1000, RRID: AB_ 2882435), anti-GLB1 (66586-1-Ig, proteintech, dilution 1:1000, RRID: AB_2881946), anti-p38 (80821-2-RR, proteintech, dilution 1:1000, RRID: AB_3670489), antip21 (ab109199, abcam, dilution 1:1000, RRID: AB_ 10861551), anti-p16 (ab189034, abcam, dilution 1:1000, RRID: RRID: AB_2737282), and anti-GAPDH (#5174, Cell Signalling Technology, dilution 1:1000, RRID: AB_10622025). ..

    Membrane:

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation.
    Article Snippet: Proteins (10–30 μg/well, equal amounts within group but different amounts among groups for better comparison) were separated on 10% or 12% SDS-PAGE gel and transferred on an equilibrated polyvinylidene difluoride membrane (PVDF, Amersham Biosciences, Buckinghamshire, 8 www.thelancet.com Vol 125 March, 2026 UK). .. After blocking with 5% bovine serum albumin (BSA, Sigma–Aldrich) in TBST (10 mM Tris–HCl, 150 mM NaCl, and 0.1% Tween 20) for 1 h at room temperature, the membrane was incubated with primary antibody overnight at 4 ◦ C. The primary antibodies contained anti-MFF (17090-1-AP, proteintech, dilution 1:1000, RRID: AB_2142463), anti-TRIM21 (67136-1-Ig, proteintech, dilution 1:1000, RRID: AB_ 2882435), anti-GLB1 (66586-1-Ig, proteintech, dilution 1:1000, RRID: AB_2881946), anti-p38 (80821-2-RR, proteintech, dilution 1:1000, RRID: AB_3670489), antip21 (ab109199, abcam, dilution 1:1000, RRID: AB_ 10861551), anti-p16 (ab189034, abcam, dilution 1:1000, RRID: RRID: AB_2737282), and anti-GAPDH (#5174, Cell Signalling Technology, dilution 1:1000, RRID: AB_10622025). ..

    Purification:

    Article Title: ACAT1/SOAT1 governs mitochondrial dynamics and thermogenesis via the mTOR-MFN2-USP2 axis in brown adipocytes
    Article Snippet: Proteins were separated by SDS-PAGE and transferred electrophoretically to a PVDF membrane (Sigma-Aldrich, 24937-79-9). .. Membranes were incubated with primary antibodies overnight at 4 °C: anti-ACAT1 (Purified by our lab), anti-HSP90 (ProteinTech, 60318-1-Ig), anti-Beta Actin (ProteinTech, 66009-1-Ig), antiOXPHOS (Abcam, ab110413), anti-UCP1 (Abcam, ab10983), antiCalnexin (ProteinTech, 10427-2-AP), anti-IP3R (ProteinTech, 19962-1- AP), anti-FACL4 (Abcam, ab155282), anti-VDAC (ProteinTech, 10866-1- AP), anti-Tubulin (ProteinTech, 66031-1-Ig), anti-MFN2 (ProteinTech, 12186-1-AP), anti-MFN1 (ProteinTech, 66776-1-Ig), anti-OPA1 (ProteinTech, 27733-1-AP), anti-DRP1 (ProteinTech, 12957-1-AP), antipSer616-DRP1 (Cell Signaling, 3455), anti-MFF (ProteinTech, 17090-1- AP), anti-Flag (Sigma, F1804), anti-P4D1 (Santa Cruz, sc-8017), anti-HA (ProteinTech, 51064-1-AP), anti-mTOR (ProteinTech, 66888-1-Ig), antipSer2448-mTOR (Cell signaling, 5536), anti-pThr389-p70S6Kinase (Cell signaling, 9234), anti-USP2 (ProteinTech, 10392-1-AP) and antiGNPAT (ProteinTech, 14931-1-AP). .. After incubation with primary antibodies, membranes were washed with TBST and incubated with secondary antibodies conjugated to horseradish peroxidase (HRP; Epizyme, LF101 and LF102) at room temperature for 1 hour.

    Cell-Signaling:

    Article Title: ACAT1/SOAT1 governs mitochondrial dynamics and thermogenesis via the mTOR-MFN2-USP2 axis in brown adipocytes
    Article Snippet: Proteins were separated by SDS-PAGE and transferred electrophoretically to a PVDF membrane (Sigma-Aldrich, 24937-79-9). .. Membranes were incubated with primary antibodies overnight at 4 °C: anti-ACAT1 (Purified by our lab), anti-HSP90 (ProteinTech, 60318-1-Ig), anti-Beta Actin (ProteinTech, 66009-1-Ig), antiOXPHOS (Abcam, ab110413), anti-UCP1 (Abcam, ab10983), antiCalnexin (ProteinTech, 10427-2-AP), anti-IP3R (ProteinTech, 19962-1- AP), anti-FACL4 (Abcam, ab155282), anti-VDAC (ProteinTech, 10866-1- AP), anti-Tubulin (ProteinTech, 66031-1-Ig), anti-MFN2 (ProteinTech, 12186-1-AP), anti-MFN1 (ProteinTech, 66776-1-Ig), anti-OPA1 (ProteinTech, 27733-1-AP), anti-DRP1 (ProteinTech, 12957-1-AP), antipSer616-DRP1 (Cell Signaling, 3455), anti-MFF (ProteinTech, 17090-1- AP), anti-Flag (Sigma, F1804), anti-P4D1 (Santa Cruz, sc-8017), anti-HA (ProteinTech, 51064-1-AP), anti-mTOR (ProteinTech, 66888-1-Ig), antipSer2448-mTOR (Cell signaling, 5536), anti-pThr389-p70S6Kinase (Cell signaling, 9234), anti-USP2 (ProteinTech, 10392-1-AP) and antiGNPAT (ProteinTech, 14931-1-AP). .. After incubation with primary antibodies, membranes were washed with TBST and incubated with secondary antibodies conjugated to horseradish peroxidase (HRP; Epizyme, LF101 and LF102) at room temperature for 1 hour.



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    Image Search Results


    List of primary antibodies, secondary antibodies, and nuclear dyes used in this study.

    Journal: Cell Death & Disease

    Article Title: Dual-faced metabolic reprogramming powers proliferation: FGF21 drives lung squamous carcinoma progression via CaMKII-DRP1-mediated mitochondrial fission

    doi: 10.1038/s41419-026-09084-3

    Figure Lengend Snippet: List of primary antibodies, secondary antibodies, and nuclear dyes used in this study.

    Article Snippet: Anti-MFF , 84580 , Rabbit , CST , WB (1:1000).

    Techniques:

    TRIM21 was an E3 ubiquitin ligase of MFF and promoted its ubiquitination degradation. (A and B) The levels of the remained MFF protein in 40 μg/mL CIP-exposed HTR-8/SVneo cells and with 10 μM CHX treatment within 10 h and its relative quantification (n = 3 independent experiments, Student's t-test). (C) The protein levels of MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells treated with 10 μM MG132 or 50 μM CQ and co-treated with 10 μM CHX and its relative quantification (n = 3 independent experiments, one-way ANOVA). (D and E) IP assay analysis of the protein levels of MFF-Ub pulled down by MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells and its relative quantification (n = 3 independent experiments, Student's t-test). (F) MFF IP-MS assay analysis of the proteins that were pulled down by MFF in HTR-8/SVneo cells. (G–K) IP assay analysis of the protein levels of TRIM21 and MFF-Ub pulled down by MFF in HTR-8/SVneo cells with overexpression (Student's t-test) or knockdown (one-way ANOVA) of TRIM21, with MFF and TRIM21 protein levels in cell lysates and their relative quantification (n = 3 independent experiments).

    Journal: eBioMedicine

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation

    doi: 10.1016/j.ebiom.2026.106146

    Figure Lengend Snippet: TRIM21 was an E3 ubiquitin ligase of MFF and promoted its ubiquitination degradation. (A and B) The levels of the remained MFF protein in 40 μg/mL CIP-exposed HTR-8/SVneo cells and with 10 μM CHX treatment within 10 h and its relative quantification (n = 3 independent experiments, Student's t-test). (C) The protein levels of MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells treated with 10 μM MG132 or 50 μM CQ and co-treated with 10 μM CHX and its relative quantification (n = 3 independent experiments, one-way ANOVA). (D and E) IP assay analysis of the protein levels of MFF-Ub pulled down by MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells and its relative quantification (n = 3 independent experiments, Student's t-test). (F) MFF IP-MS assay analysis of the proteins that were pulled down by MFF in HTR-8/SVneo cells. (G–K) IP assay analysis of the protein levels of TRIM21 and MFF-Ub pulled down by MFF in HTR-8/SVneo cells with overexpression (Student's t-test) or knockdown (one-way ANOVA) of TRIM21, with MFF and TRIM21 protein levels in cell lysates and their relative quantification (n = 3 independent experiments).

    Article Snippet: The lysates were incubated with MFF antibody (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463 ) or TRIM21 antibody (67136-1-Ig, proteintech, dilution 1:200, RRID: AB_2882435 ) at 4 °C overnight, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931 ) as negative control, and then incubated with Protein A/G magnetic beads (HY–K0202, MedChemExpress) for another 6 h to form bead-immunoprecipitate complex.

    Techniques: Ubiquitin Proteomics, Quantitative Proteomics, Protein-Protein interactions, Over Expression, Knockdown

    The joint roles of CIP and TRIM21 in MFF ubiquitination degradation. (A) TRIM21 mRNA levels in 0, 1, 10, 20, or 40 μg/mL CIP-exposed HTR-8/SVneo cells (n = 3 independent experiments, one-way ANOVA). (B) TRIM21 protein levels in 0, 1, 10, 20, or 40 μg/mL CIP-exposed HTR-8/SVneo cells and its relative quantification (n = 3 independent experiments, one-way ANOVA). (C and D) IP assay analysis of the protein levels of MFF pulled down by TRIM21 in 40 μg/mL CIP-exposed HTR-8/SVneo cells with MFF and TRIM21 protein levels in cell lysates and its relative quantification (n = 3 independent experiments, Student's t-test). (E–I) IP assay analysis of the protein levels of TRIM21 and MFF-Ub pulled down by MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells with TRIM21 knockdown, with MFF and TRIM21 protein levels in cell lysates and their relative quantification (n = 3 independent experiments, one-way ANOVA).

    Journal: eBioMedicine

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation

    doi: 10.1016/j.ebiom.2026.106146

    Figure Lengend Snippet: The joint roles of CIP and TRIM21 in MFF ubiquitination degradation. (A) TRIM21 mRNA levels in 0, 1, 10, 20, or 40 μg/mL CIP-exposed HTR-8/SVneo cells (n = 3 independent experiments, one-way ANOVA). (B) TRIM21 protein levels in 0, 1, 10, 20, or 40 μg/mL CIP-exposed HTR-8/SVneo cells and its relative quantification (n = 3 independent experiments, one-way ANOVA). (C and D) IP assay analysis of the protein levels of MFF pulled down by TRIM21 in 40 μg/mL CIP-exposed HTR-8/SVneo cells with MFF and TRIM21 protein levels in cell lysates and its relative quantification (n = 3 independent experiments, Student's t-test). (E–I) IP assay analysis of the protein levels of TRIM21 and MFF-Ub pulled down by MFF in 40 μg/mL CIP-exposed HTR-8/SVneo cells with TRIM21 knockdown, with MFF and TRIM21 protein levels in cell lysates and their relative quantification (n = 3 independent experiments, one-way ANOVA).

    Article Snippet: The lysates were incubated with MFF antibody (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463 ) or TRIM21 antibody (67136-1-Ig, proteintech, dilution 1:200, RRID: AB_2882435 ) at 4 °C overnight, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931 ) as negative control, and then incubated with Protein A/G magnetic beads (HY–K0202, MedChemExpress) for another 6 h to form bead-immunoprecipitate complex.

    Techniques: Ubiquitin Proteomics, Quantitative Proteomics, Knockdown

    Verification of TRIM21/MFF signalling in human villous tissues. (A and B) TRIM21 protein levels in HC and UM villous tissues and its relative quantification (n = 12, Student's t-test). (C–G) IP assay analysis of the protein levels of TRIM21 and MFF-Ub that were pulled down by MFF in HC and UM villous tissues, with MFF and TRIM21 protein levels in tissue lysates and their relative quantification (each n = 12, Student's t-test). (H) Multivariate logistic regression analysis of forest plot of odds ratio (OR) of TRIM21 protein levels in villous tissues with its 95% confidence interval (95% CI) (n = 12).

    Journal: eBioMedicine

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation

    doi: 10.1016/j.ebiom.2026.106146

    Figure Lengend Snippet: Verification of TRIM21/MFF signalling in human villous tissues. (A and B) TRIM21 protein levels in HC and UM villous tissues and its relative quantification (n = 12, Student's t-test). (C–G) IP assay analysis of the protein levels of TRIM21 and MFF-Ub that were pulled down by MFF in HC and UM villous tissues, with MFF and TRIM21 protein levels in tissue lysates and their relative quantification (each n = 12, Student's t-test). (H) Multivariate logistic regression analysis of forest plot of odds ratio (OR) of TRIM21 protein levels in villous tissues with its 95% confidence interval (95% CI) (n = 12).

    Article Snippet: The lysates were incubated with MFF antibody (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463 ) or TRIM21 antibody (67136-1-Ig, proteintech, dilution 1:200, RRID: AB_2882435 ) at 4 °C overnight, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931 ) as negative control, and then incubated with Protein A/G magnetic beads (HY–K0202, MedChemExpress) for another 6 h to form bead-immunoprecipitate complex.

    Techniques: Quantitative Proteomics

    Verification of murine Trim21/Mff signalling in CIP-exposed mouse placental tissues. (A) Murine Trim21 mRNA levels in placental tissues of CIP-exposed mice (n = 6, one-way ANOVA). (B and C) Murine Trim21 protein levels in placental tissues of CIP-exposed mice and its relative quantification (n = 6, one-way ANOVA). (D) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in placental tissues of 160 mg/kg/d CIP-exposed mice, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (E) Schematic diagram of CIP-exposed mouse model with Trim21 knockdown. Pregnant mice were treated with saline or 160 mg/kg/d CIP and also treated with 10 mg/kg/3 d AS-Trim 21 (with AS–NC as control). (F and G) Embryo resorption (indicated by red arrows) and the average miscarriage rates in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6, one-way ANOVA). (H) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (I) The quantification of positive intensity of SA-β-Galactose staining of placental tissues in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6, one-way ANOVA). (J) The protein levels of p16, p21, p38, and β-gal in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6). (K) The relative mtDNA copy number in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA). (L) The NAD + /NADH ratios in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA). (M) The MMP levels in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA).

    Journal: eBioMedicine

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation

    doi: 10.1016/j.ebiom.2026.106146

    Figure Lengend Snippet: Verification of murine Trim21/Mff signalling in CIP-exposed mouse placental tissues. (A) Murine Trim21 mRNA levels in placental tissues of CIP-exposed mice (n = 6, one-way ANOVA). (B and C) Murine Trim21 protein levels in placental tissues of CIP-exposed mice and its relative quantification (n = 6, one-way ANOVA). (D) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in placental tissues of 160 mg/kg/d CIP-exposed mice, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (E) Schematic diagram of CIP-exposed mouse model with Trim21 knockdown. Pregnant mice were treated with saline or 160 mg/kg/d CIP and also treated with 10 mg/kg/3 d AS-Trim 21 (with AS–NC as control). (F and G) Embryo resorption (indicated by red arrows) and the average miscarriage rates in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6, one-way ANOVA). (H) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (I) The quantification of positive intensity of SA-β-Galactose staining of placental tissues in 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6, one-way ANOVA). (J) The protein levels of p16, p21, p38, and β-gal in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (each n = 6). (K) The relative mtDNA copy number in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA). (L) The NAD + /NADH ratios in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA). (M) The MMP levels in placental tissues of 160 mg/kg/d CIP-exposed mice with Trim21 knockdown (n = 6, one-way ANOVA).

    Article Snippet: The lysates were incubated with MFF antibody (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463 ) or TRIM21 antibody (67136-1-Ig, proteintech, dilution 1:200, RRID: AB_2882435 ) at 4 °C overnight, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931 ) as negative control, and then incubated with Protein A/G magnetic beads (HY–K0202, MedChemExpress) for another 6 h to form bead-immunoprecipitate complex.

    Techniques: Quantitative Proteomics, Knockdown, Saline, Control, Staining

    Miscarriage treatment. (A) Schematic diagram of CIP-exposed mouse model with F + Q treatment. Pregnant mice were treated with saline or 160 mg/kg/d CIP and also treated with 50 mg/kg/2 d F and 10 mg/kg/2 d Q. (B–D) The protein levels of Trim21 and Mff in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment and their relative quantification (each n = 6, Student's t-test). (E and F) Embryo resorption (indicated by red arrows) and the average miscarriage rates in 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6, Student's t-test). (G) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in 160 mg/kg/d CIP-exposed mice with F + Q treatment, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (H) The quantification of positive intensity of SA-β-Galactose staining of placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6, Student's t-test). (I) The protein levels of p16, p21, p38, and β-gal in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6). (J) The relative mtDNA copy number in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (K) The NAD + /NADH ratios in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (L) The MMP levels in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (M) The proposed mechanisms.

    Journal: eBioMedicine

    Article Title: Environmental ciprofloxacin triggers pregnancy loss: senescence-driven miscarriage via TRIM21-mediated MFF degradation

    doi: 10.1016/j.ebiom.2026.106146

    Figure Lengend Snippet: Miscarriage treatment. (A) Schematic diagram of CIP-exposed mouse model with F + Q treatment. Pregnant mice were treated with saline or 160 mg/kg/d CIP and also treated with 50 mg/kg/2 d F and 10 mg/kg/2 d Q. (B–D) The protein levels of Trim21 and Mff in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment and their relative quantification (each n = 6, Student's t-test). (E and F) Embryo resorption (indicated by red arrows) and the average miscarriage rates in 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6, Student's t-test). (G) IP assay analysis of the protein levels of Trim21 and Mff-Ub that were pulled down by Mff in 160 mg/kg/d CIP-exposed mice with F + Q treatment, with Mff and Trim21 protein levels in tissue lysates (each n = 6). (H) The quantification of positive intensity of SA-β-Galactose staining of placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6, Student's t-test). (I) The protein levels of p16, p21, p38, and β-gal in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (each n = 6). (J) The relative mtDNA copy number in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (K) The NAD + /NADH ratios in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (L) The MMP levels in placental tissues of 160 mg/kg/d CIP-exposed mice with F + Q treatment (n = 6, Student's t-test). (M) The proposed mechanisms.

    Article Snippet: The lysates were incubated with MFF antibody (17090-1-AP, proteintech, dilution 1:200, RRID: AB_2142463 ) or TRIM21 antibody (67136-1-Ig, proteintech, dilution 1:200, RRID: AB_2882435 ) at 4 °C overnight, with equal weight of IgG antibody (ab172730, Abcam, dilution 1:200, RRID: AB_2687931 ) as negative control, and then incubated with Protein A/G magnetic beads (HY–K0202, MedChemExpress) for another 6 h to form bead-immunoprecipitate complex.

    Techniques: Saline, Quantitative Proteomics, Staining