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anti resource source identifier rabbit polyclonal fancd2 novus biologicals nb100-182  (Novus Biologicals)


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    Novus Biologicals anti resource source identifier rabbit polyclonal fancd2 novus biologicals nb100-182
    Anti Resource Source Identifier Rabbit Polyclonal Fancd2 Novus Biologicals Nb100 182, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 95/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/fancd2/FANCD2+Antibody+-+BSA+Free/pm37601968-256-2-9
    Average 95 stars, based on 188 article reviews
    anti resource source identifier rabbit polyclonal fancd2 novus biologicals nb100-182 - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: SMAD4 loss drives cancer chromosomal instability through transcription-replication conflict-induced replication stress
    Article Snippet: To detect S-phase cells, cells were labeled with 20 μM EdU (Jena Biosciences, Germany) for 30 minutes and EdU incorporation was detected using the Click-iT EdU Cell Proliferation Kit for Imaging with Alexa Fluor 488 dye according to the manufactureŕs recommendation (C10337; Invitrogen, USA). .. After blocking, cells were incubated overnight at 4°C with the following primary antibodies (diluted 1:750 in 3% BSA in PBS): anti-53BP1 (H-300; Santa Cruz, USA; cat no sc-22760; RRID: AB_2256326), anti-FANCD2 (Novus Biologicals, USA; cat no NB100-182; RRID: AB_3149940), anti-Phospho H2A.X (Ser139; ψ-H2AX) (JBW301; mouse; Merck Millipore, Germany; cat no #05-636; RRID: AB_309864), anti-CENP-C (MBL International, USA; cat no PD030; RRID: AB_10693556). .. This was followed by incubation with Alexa Fluor®-conjugated secondary antibodies (1:1000; Alexa Fluor 488, Alexa Fluor 594 and Alexa Fluor 647; Thermo Fisher Scientific, USA; cat no A-11029, A-11076, A-11034, A-11012 and A-21236) for 1 hour at room temperature.

    Incubation:

    Article Title: SMAD4 loss drives cancer chromosomal instability through transcription-replication conflict-induced replication stress
    Article Snippet: To detect S-phase cells, cells were labeled with 20 μM EdU (Jena Biosciences, Germany) for 30 minutes and EdU incorporation was detected using the Click-iT EdU Cell Proliferation Kit for Imaging with Alexa Fluor 488 dye according to the manufactureŕs recommendation (C10337; Invitrogen, USA). .. After blocking, cells were incubated overnight at 4°C with the following primary antibodies (diluted 1:750 in 3% BSA in PBS): anti-53BP1 (H-300; Santa Cruz, USA; cat no sc-22760; RRID: AB_2256326), anti-FANCD2 (Novus Biologicals, USA; cat no NB100-182; RRID: AB_3149940), anti-Phospho H2A.X (Ser139; ψ-H2AX) (JBW301; mouse; Merck Millipore, Germany; cat no #05-636; RRID: AB_309864), anti-CENP-C (MBL International, USA; cat no PD030; RRID: AB_10693556). .. This was followed by incubation with Alexa Fluor®-conjugated secondary antibodies (1:1000; Alexa Fluor 488, Alexa Fluor 594 and Alexa Fluor 647; Thermo Fisher Scientific, USA; cat no A-11029, A-11076, A-11034, A-11012 and A-21236) for 1 hour at room temperature.

    Article Title: Aurora kinase A is a synthetic lethal target in FANCA-deficient cancers
    Article Snippet: .. Cells were then incubated with the primary antibody for 1 hour at RT, rabbit anti-FANCD2 antibody (Novus Biologicals, NB100-182, 1:500 in SB) or mouse anti-FANCD2 antibody (Novus Biologicals, NB100-316, 1:500 in SB). .. After 2 washes with SB, cells were incubated with goat anti-rabbit Alexa Fluor 647 (Jackson ImmunoResearch, 1:2500 in SB) for 1 hour at RT.

    Cell-Signaling:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Control:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Western Blot:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Immunofluorescence:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Immunohistochemistry:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Staining:

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD
    Article Snippet: The commercial sources of major reagents and chemicals include: Mitomycin C (MMC, S8146, Selleck, Shanghai, China), Cisplatin (CDDP, MB1055, Meilunbio, Dalian, China), SU5416 (HY‐10374, MedChemExpress, Monmouth Junction, NJ, USA), Amifostine trihydrate (S4298, Selleck, Shanghai, China). .. Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining. ..

    Proximity Ligation Assay:

    Article Title: FANCD2 restrains fork progression and prevents fragility at early origins upon re-replication
    Article Snippet: .. The PLA reaction was performed using the following reagents: PCNA (PC10, sc-56, Santa Cruz), MCM2 (D7G11, 3619, Cell Signaling), FANCD2 (NB100-182, Novus Biologicals), anti-Biotin (200-002-211, Jackson ImmunoResearch), Duolink in situ PLA probe anti-rabbit PLUS (Merck, DUO92002), Duolink in situ PLA probe anti-mouse MINUS (Merck, DUO92004) and Duolink-Detection Reagents Red (Merck). .. Finally, nuclei were stained with DAPI and mounted with MOWIOL® 4–88. (Sigma-Aldrich).

    In Situ:

    Article Title: FANCD2 restrains fork progression and prevents fragility at early origins upon re-replication
    Article Snippet: .. The PLA reaction was performed using the following reagents: PCNA (PC10, sc-56, Santa Cruz), MCM2 (D7G11, 3619, Cell Signaling), FANCD2 (NB100-182, Novus Biologicals), anti-Biotin (200-002-211, Jackson ImmunoResearch), Duolink in situ PLA probe anti-rabbit PLUS (Merck, DUO92002), Duolink in situ PLA probe anti-mouse MINUS (Merck, DUO92004) and Duolink-Detection Reagents Red (Merck). .. Finally, nuclei were stained with DAPI and mounted with MOWIOL® 4–88. (Sigma-Aldrich).

    other:

    Article Title: EP300 deficiency leads to chronic replication stress mediated by defective replication fork protection
    Article Snippet: Images were captured using- 63 mm oil immersion lens, Zeiss Axio microscope.

    Article Title: CTCF/cohesin-binding sites are susceptible to replication-associated DNA damage and genomic instability in cancer cells
    Article Snippet: Anti-FANCD2, rabbit polyclonal , Novus , Cat# NB 100–182; RRID: AB_350110.



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    Santa Cruz Biotechnology fancd2
    Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, <t>FANCD2,</t> RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.
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    Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, <t>FANCD2,</t> RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.
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    Image Search Results


    MMC or CDDP Treatment Induces Progressive DNA Damage Accumulation in Pulmonary Vascular Endothelium. (A–H) Immunoblots and analyzed graphs showing the γ‐H2AX protein expression in the lung tissue of rats treated with MMC (3 mg/kg, once weekly) for 2 or 4 weeks (A–D), and CDDP (4 mg/kg, once weekly) for 2 or 4 weeks (E–H). n = 6–12 rats per group. Protein expression levels were calculated as the ratio of the target protein band intensity to that of β‑actin and were normalized to the mean values of the respective 2‑week or 4‑week NC groups, expressed as fold change. (I, J) Immunohistochemical staining for the FA core factors (FANCL and FANCD2) and the DNA damage marker γ‐H2AX in lung sections from rats treated with MMC (3 mg/kg, once weekly) or CDDP (4 mg/kg, once weekly) for 2 or 4 weeks, followed by quantification of the percentage of positive cells within the pulmonary vasculature. n = 6 rats per group. (K, L) Immunofluorescence staining for the endothelial cell marker vWF and the DNA damage marker γ‐H2AX in lung sections from rats treated with MMC (3 mg/kg, once weekly) or CDDP (4 mg/kg, once weekly) for 4 weeks, followed by correlation analysis between γ‐H2AX fluorescence intensity within vWF‑positive regions and vessel wall area percentage. n = 6 rats per group, with 10–25 vessels analyzed per group. Data are presented as mean ± SD. Differences between two groups were analyzed by t ‐test for C (FANCD2, FANCL), D (all panels), G (FANCD2, γ‐H2AX), H (all panels), and L (CDDP), Mann‐Whitney U‐test for C (γ‐H2AX), D (PCNA), G (FANCL), and L (MMC), or one‐way ANOVA with Tukey's honestly significant difference (Tukey's HSD) post hoc test for J. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in I and 25 µm in K. NC: normal control.

    Journal: Advanced Science

    Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD

    doi: 10.1002/advs.78081

    Figure Lengend Snippet: MMC or CDDP Treatment Induces Progressive DNA Damage Accumulation in Pulmonary Vascular Endothelium. (A–H) Immunoblots and analyzed graphs showing the γ‐H2AX protein expression in the lung tissue of rats treated with MMC (3 mg/kg, once weekly) for 2 or 4 weeks (A–D), and CDDP (4 mg/kg, once weekly) for 2 or 4 weeks (E–H). n = 6–12 rats per group. Protein expression levels were calculated as the ratio of the target protein band intensity to that of β‑actin and were normalized to the mean values of the respective 2‑week or 4‑week NC groups, expressed as fold change. (I, J) Immunohistochemical staining for the FA core factors (FANCL and FANCD2) and the DNA damage marker γ‐H2AX in lung sections from rats treated with MMC (3 mg/kg, once weekly) or CDDP (4 mg/kg, once weekly) for 2 or 4 weeks, followed by quantification of the percentage of positive cells within the pulmonary vasculature. n = 6 rats per group. (K, L) Immunofluorescence staining for the endothelial cell marker vWF and the DNA damage marker γ‐H2AX in lung sections from rats treated with MMC (3 mg/kg, once weekly) or CDDP (4 mg/kg, once weekly) for 4 weeks, followed by correlation analysis between γ‐H2AX fluorescence intensity within vWF‑positive regions and vessel wall area percentage. n = 6 rats per group, with 10–25 vessels analyzed per group. Data are presented as mean ± SD. Differences between two groups were analyzed by t ‐test for C (FANCD2, FANCL), D (all panels), G (FANCD2, γ‐H2AX), H (all panels), and L (CDDP), Mann‐Whitney U‐test for C (γ‐H2AX), D (PCNA), G (FANCL), and L (MMC), or one‐way ANOVA with Tukey's honestly significant difference (Tukey's HSD) post hoc test for J. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in I and 25 µm in K. NC: normal control.

    Article Snippet: Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining.

    Techniques: Western Blot, Expressing, Tissue, Immunohistochemical staining, Staining, Marker, Immunofluorescence, Analysis, Fluorescence, MANN-WHITNEY, Control

    Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, FANCD2, RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

    Journal: Blood Advances

    Article Title: Inhibition of NAMPT targets DNA damage response to sensitize alkylating chemotherapy in TP53 mutant mantle cell lymphoma ∗

    doi: 10.1182/bloodadvances.2025016765

    Figure Lengend Snippet: Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, FANCD2, RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

    Article Snippet: Western blotting (WB) was performed to evaluate the expression levels of total protein and phospho-specific isoforms using the following antibodies: FANCD2 (Santa Cruz Biotechnology, sc-20022), RAD51 (Santa Cruz Biotechnology, sc-398587), PBEF (Santa Cruz Biotechnology, sc-166946), cleaved Caspase3 (Cell Signaling Technology, 9664s), PARP (Cell Signaling Technology, 9532s), cleaved PARP (Cell Signaling Technology, 5625s), γ-H2AX (Ser139; Cell Signaling Technology, 9718s), p-CHK1 (Ser345; Cell Signaling Technology, 2341s), p53 (Cell Signaling Technology, 2527s), p-ATR (Ser428; Cell Signaling Technology, 2853s), p-ATM (Ser1981; Cell Signaling Technology, 4526s), and p-CHK2 (Thr68; Cell Signaling Technology, 2197s).

    Techniques: Inhibition, In Vivo, Injection, Radio Immunoprecipitation, Control

    Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, FANCD2, RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

    Journal: Blood Advances

    Article Title: Inhibition of NAMPT targets DNA damage response to sensitize alkylating chemotherapy in TP53 mutant mantle cell lymphoma ∗

    doi: 10.1182/bloodadvances.2025016765

    Figure Lengend Snippet: Inhibition of NAMPT function via KPT-9274 inhibits tumor growth in vivo. (A) NOG mice were injected subcutaneously with Mino cells. After detection of the tumor, the mice were randomized and treated orally with either KPT-9274 or the vehicle for 5 consecutive days per week for 3 weeks. Tumor volume was evaluated via caliper measurement. Differences between the 2 groups were evaluated using the standard t test. (B) Tumor cells collected from mice were lysed in radio-immunoprecipitation assay buffer, and the whole-cell lysate was subjected to WB analysis and probed with antibodies against Cl-PARP, FANCD2, RAD51, and γ-H2AX. GAPDH was used as the loading control. ns > .05; ∗∗ P ≤ .01; ∗∗∗ P ≤ .001; ∗∗∗∗ P ≤ .0001.

    Article Snippet: Western blotting (WB) was performed to evaluate the expression levels of total protein and phospho-specific isoforms using the following antibodies: FANCD2 (Santa Cruz Biotechnology, sc-20022), RAD51 (Santa Cruz Biotechnology, sc-398587), PBEF (Santa Cruz Biotechnology, sc-166946), cleaved Caspase3 (Cell Signaling Technology, 9664s), PARP (Cell Signaling Technology, 9532s), cleaved PARP (Cell Signaling Technology, 5625s), γ-H2AX (Ser139; Cell Signaling Technology, 9718s), p-CHK1 (Ser345; Cell Signaling Technology, 2341s), p53 (Cell Signaling Technology, 2527s), p-ATR (Ser428; Cell Signaling Technology, 2853s), p-ATM (Ser1981; Cell Signaling Technology, 4526s), and p-CHK2 (Thr68; Cell Signaling Technology, 2197s).

    Techniques: Inhibition, In Vivo, Injection, Radio Immunoprecipitation, Control