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polyclonal anti rev7 mad2l2  (Proteintech)


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    Proteintech polyclonal anti rev7 mad2l2
    Polyclonal Anti Rev7 Mad2l2, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/12683+1+ap/pmc12820532-83-12-23?v=Proteintech
    Average 93 stars, based on 19 article reviews
    polyclonal anti rev7 mad2l2 - by Bioz Stars, 2026-07
    93/100 stars

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    polyclonal anti rev7 mad2l2  (Proteintech)
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    H1299 WT and RNF25 KO cells were transfected with indicated siRNAs 48 h before conducting the DNA fork movement assay ( a ) and the fork degradation assay ( b ). 150 fibers were analyzed per condition with means indicated with bars. Each Fig. shown is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c HA-RNF25 WT and domain mutants were expressed in H1299 cells using adenoviral vectors, then immunoprecipitated with anti-HA beads. d SIRF assay between HA-RNF25 (WT and C135/138S) and EdU in H1299 RNF25 KO cells transfected with control or <t>REV7</t> siRNA. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in purple and blue, with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-purple, replicate 2-blue): (99, 64); (75, 96); (77, 75); (84, 74); (76, 82); (78, 61); (88, 86); (87, 80); (75, 70); (81, 66); (76, 77); (77, 63). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. e SIRF assay between HA-REV7 (WT and ΔSB) and EdU in H1299 WT and RNF25 KO cells. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in blue and pink with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-blue, replicate 2-pink): (70, 75); (74, 77); (71, 87); (70, 76); (73, 85); (72, 76); (70, 76); (75, 86); (72, 83); (74, 75); (65, 80); (72, 79). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. f Heatmap depicting relative mRNA expression of replication and DNA damage repair factors in PDACs from TCGA. Samples were clustered as normal or tumor samples, with additional classification of tumor samples by mutation status of key PDAC driver genes and genome maintenance related features (RS – Replication Stress signature, HRD – Homologous Recombination Deficiency score, CA20 – Centrosome Amplification signature). Source data are provided as a file.
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    H1299 WT and RNF25 KO cells were transfected with indicated siRNAs 48 h before conducting the DNA fork movement assay ( a ) and the fork degradation assay ( b ). 150 fibers were analyzed per condition with means indicated with bars. Each Fig. shown is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c HA-RNF25 WT and domain mutants were expressed in H1299 cells using adenoviral vectors, then immunoprecipitated with anti-HA beads. d SIRF assay between HA-RNF25 (WT and C135/138S) and EdU in H1299 RNF25 KO cells transfected with control or <t>REV7</t> siRNA. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in purple and blue, with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-purple, replicate 2-blue): (99, 64); (75, 96); (77, 75); (84, 74); (76, 82); (78, 61); (88, 86); (87, 80); (75, 70); (81, 66); (76, 77); (77, 63). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. e SIRF assay between HA-REV7 (WT and ΔSB) and EdU in H1299 WT and RNF25 KO cells. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in blue and pink with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-blue, replicate 2-pink): (70, 75); (74, 77); (71, 87); (70, 76); (73, 85); (72, 76); (70, 76); (75, 86); (72, 83); (74, 75); (65, 80); (72, 79). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. f Heatmap depicting relative mRNA expression of replication and DNA damage repair factors in PDACs from TCGA. Samples were clustered as normal or tumor samples, with additional classification of tumor samples by mutation status of key PDAC driver genes and genome maintenance related features (RS – Replication Stress signature, HRD – Homologous Recombination Deficiency score, CA20 – Centrosome Amplification signature). Source data are provided as a file.
    Rabbit Polyclonal Anti Mad2l2 Rev7 Proteintech, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ADAM9 induces pyroptosis <t>in</t> <t>GHRH</t> neurons in SAH. ( A ) The content of ADAM9 in the CSF of mice on day 7 after SAH surgery was significantly greater than that in the CSF of sham-operated mice. n = 3. ( B ) The expression of ADAM9 and the activation of microglia at 3 d, 7 d and 14 d after SAH surgery compared with those in sham‐operated mice. Iba-1 is a microglial marker. Scale bar = 100 μm. ( C ) Quantification of B. n = 3. ( D )The content of ADAM9 in the supernatant of GHRH neurons. Microglial supernatant was used as a positive control. n = 3. ( E ) CD11b and Iba-1 were coexpressed in the SAH group, and microglia were more strongly activated in the SAH group than in the sham‐operated group. CD11b is an M1 microglial marker. The arrow indicates microglia. Scale bar = 20 μm. ( F ) ADAM9 and Iba-1 were coexpressed in hypothalamic microglia in primary culture on day 7 after SAH surgery. Scale bar = 20 μm. ( G ) Caspase-1 was highly expressed in the ARC at 7 days after SAH surgery. GAPDH expression was used to confirm equal protein loading and blotting. ( H ) Quantification of G. n = 3. ( I ) Immunofluorescence showed that the number of GHRH + <t>GSDMD</t> + neurons in the SAH group was significantly greater than that in the control group. ( J ) Quantification of the ARC in I. n = 3. ( K ) The expression of IL-1β, IL-6 and IL-8 in the ARC on day 7 after SAH surgery was significantly higher than that in the sham‐operated mice. n = 3. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; *** p < 0.001
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    H1299 WT and RNF25 KO cells were transfected with indicated siRNAs 48 h before conducting the DNA fork movement assay ( a ) and the fork degradation assay ( b ). 150 fibers were analyzed per condition with means indicated with bars. Each Fig. shown is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c HA-RNF25 WT and domain mutants were expressed in H1299 cells using adenoviral vectors, then immunoprecipitated with anti-HA beads. d SIRF assay between HA-RNF25 (WT and C135/138S) and EdU in H1299 RNF25 KO cells transfected with control or REV7 siRNA. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in purple and blue, with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-purple, replicate 2-blue): (99, 64); (75, 96); (77, 75); (84, 74); (76, 82); (78, 61); (88, 86); (87, 80); (75, 70); (81, 66); (76, 77); (77, 63). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. e SIRF assay between HA-REV7 (WT and ΔSB) and EdU in H1299 WT and RNF25 KO cells. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in blue and pink with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-blue, replicate 2-pink): (70, 75); (74, 77); (71, 87); (70, 76); (73, 85); (72, 76); (70, 76); (75, 86); (72, 83); (74, 75); (65, 80); (72, 79). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. f Heatmap depicting relative mRNA expression of replication and DNA damage repair factors in PDACs from TCGA. Samples were clustered as normal or tumor samples, with additional classification of tumor samples by mutation status of key PDAC driver genes and genome maintenance related features (RS – Replication Stress signature, HRD – Homologous Recombination Deficiency score, CA20 – Centrosome Amplification signature). Source data are provided as a file.

    Journal: Nature Communications

    Article Title: The RING finger E3 ligase RNF25 protects DNA replication forks independently of its canonical roles in ubiquitin signaling

    doi: 10.1038/s41467-025-62368-8

    Figure Lengend Snippet: H1299 WT and RNF25 KO cells were transfected with indicated siRNAs 48 h before conducting the DNA fork movement assay ( a ) and the fork degradation assay ( b ). 150 fibers were analyzed per condition with means indicated with bars. Each Fig. shown is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c HA-RNF25 WT and domain mutants were expressed in H1299 cells using adenoviral vectors, then immunoprecipitated with anti-HA beads. d SIRF assay between HA-RNF25 (WT and C135/138S) and EdU in H1299 RNF25 KO cells transfected with control or REV7 siRNA. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in purple and blue, with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-purple, replicate 2-blue): (99, 64); (75, 96); (77, 75); (84, 74); (76, 82); (78, 61); (88, 86); (87, 80); (75, 70); (81, 66); (76, 77); (77, 63). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. e SIRF assay between HA-REV7 (WT and ΔSB) and EdU in H1299 WT and RNF25 KO cells. The number of foci per nucleus was quantified using ImageJ. Each biological replicate is shown in blue and pink with means indicated using bars. Number of nuclei quantified per condition (from left to right) (replicate 1-blue, replicate 2-pink): (70, 75); (74, 77); (71, 87); (70, 76); (73, 85); (72, 76); (70, 76); (75, 86); (72, 83); (74, 75); (65, 80); (72, 79). Statistics: one-way ANOVA with Tukey’s multiple comparisons test on Log 2 -transformed data. f Heatmap depicting relative mRNA expression of replication and DNA damage repair factors in PDACs from TCGA. Samples were clustered as normal or tumor samples, with additional classification of tumor samples by mutation status of key PDAC driver genes and genome maintenance related features (RS – Replication Stress signature, HRD – Homologous Recombination Deficiency score, CA20 – Centrosome Amplification signature). Source data are provided as a file.

    Article Snippet: Antibodies used: TRIM11 (ab111694), pIRF3 S386 (ab76493), MUS81 (ab14387), and MRE11 (ab214) from Abcam; RNF25 (A303-844A), pRPA S33 (A300-246A), pRPA S4/S8 (A300-245A), HLTF (A300-230A), Pol η (A301-231A), Pol κ (A301-975A), Pol ι (A301-304A), RAD18 (A301-340A), and HA (A190-138A) from Bethyl Laboratories; Vinculin (V4505), SHLD3 (HPA068764), and γH2AX (05-636) from Sigma Millipore; RPA 34 (NA19L) from Calbiochem; pATM S1981 (sc-47739), β-actin (sc-47778), GAPDH (sc-32233), TNKS1/2 (sc-365897), PARP1 (sc-8007), UBE2D2 (sc-100617), REV1 (sc-393022), FANCD2 (sc-28194), Histone H2B (sc-10808), WEE1 (sc-9037), CDK2 pT14/Y15 (sc-28435-R), CDK2 (sc-6248), and PCNA (sc-56) from Santa Cruz Biotech; CHK1 (CST 2360), CHK1 pS317 (CST 2344), CDC2 pTyr15 (CST 9111), PIAS1 (CST 3550), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (CST 9106), pSTING Ser366 (CST 19781), STING (CST 13647), pSTAT1 Tyr701 (CST 9167) and STAT1 (CST 9175) from Cell Signaling Technology; REV7 (12683-1-AP), EXO1 (16253-1-AP) from Proteintech; CtIP (61141) from Active Motif.

    Techniques: Transfection, Motility Assay, Degradation Assay, Two Tailed Test, MANN-WHITNEY, Immunoprecipitation, Control, Transformation Assay, Expressing, Mutagenesis, Homologous Recombination, Amplification

    H1299 WT and RNF25 KO cells were transfected with indicated siRNAs for 48 h before conducting the DNA fiber fork degradation ( a ) or fork movement ( b ) assay. 150 fibers were analyzed per condition with means shown using bars. This Fig. is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c H1299 cells were transfected with indicated siRNAs before HA-RNF25 was expressed using adenoviral vectors and immunoprecipitated with anti-HA beads. d Schematic depicting the role of RNF25 in basal replication and following replication stress. Under basal conditions, RNF25 promotes fork movement along with REV7 and REV3L. Under replication stress conditions, RNF25 protects reversed forks by recruiting REV7. REV1 and REV3L also function in the same fork protection pathway. However, in RNF25 deficient cells, fork degradation is mediated by nucleases MRE11 and CtIP. Source data are provided as a file.

    Journal: Nature Communications

    Article Title: The RING finger E3 ligase RNF25 protects DNA replication forks independently of its canonical roles in ubiquitin signaling

    doi: 10.1038/s41467-025-62368-8

    Figure Lengend Snippet: H1299 WT and RNF25 KO cells were transfected with indicated siRNAs for 48 h before conducting the DNA fiber fork degradation ( a ) or fork movement ( b ) assay. 150 fibers were analyzed per condition with means shown using bars. This Fig. is a representative experiment from two biological replicates. Statistics: two-tailed Mann-Whitney test. c H1299 cells were transfected with indicated siRNAs before HA-RNF25 was expressed using adenoviral vectors and immunoprecipitated with anti-HA beads. d Schematic depicting the role of RNF25 in basal replication and following replication stress. Under basal conditions, RNF25 promotes fork movement along with REV7 and REV3L. Under replication stress conditions, RNF25 protects reversed forks by recruiting REV7. REV1 and REV3L also function in the same fork protection pathway. However, in RNF25 deficient cells, fork degradation is mediated by nucleases MRE11 and CtIP. Source data are provided as a file.

    Article Snippet: Antibodies used: TRIM11 (ab111694), pIRF3 S386 (ab76493), MUS81 (ab14387), and MRE11 (ab214) from Abcam; RNF25 (A303-844A), pRPA S33 (A300-246A), pRPA S4/S8 (A300-245A), HLTF (A300-230A), Pol η (A301-231A), Pol κ (A301-975A), Pol ι (A301-304A), RAD18 (A301-340A), and HA (A190-138A) from Bethyl Laboratories; Vinculin (V4505), SHLD3 (HPA068764), and γH2AX (05-636) from Sigma Millipore; RPA 34 (NA19L) from Calbiochem; pATM S1981 (sc-47739), β-actin (sc-47778), GAPDH (sc-32233), TNKS1/2 (sc-365897), PARP1 (sc-8007), UBE2D2 (sc-100617), REV1 (sc-393022), FANCD2 (sc-28194), Histone H2B (sc-10808), WEE1 (sc-9037), CDK2 pT14/Y15 (sc-28435-R), CDK2 (sc-6248), and PCNA (sc-56) from Santa Cruz Biotech; CHK1 (CST 2360), CHK1 pS317 (CST 2344), CDC2 pTyr15 (CST 9111), PIAS1 (CST 3550), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (CST 9106), pSTING Ser366 (CST 19781), STING (CST 13647), pSTAT1 Tyr701 (CST 9167) and STAT1 (CST 9175) from Cell Signaling Technology; REV7 (12683-1-AP), EXO1 (16253-1-AP) from Proteintech; CtIP (61141) from Active Motif.

    Techniques: Transfection, Two Tailed Test, MANN-WHITNEY, Immunoprecipitation

    ADAM9 induces pyroptosis in GHRH neurons in SAH. ( A ) The content of ADAM9 in the CSF of mice on day 7 after SAH surgery was significantly greater than that in the CSF of sham-operated mice. n = 3. ( B ) The expression of ADAM9 and the activation of microglia at 3 d, 7 d and 14 d after SAH surgery compared with those in sham‐operated mice. Iba-1 is a microglial marker. Scale bar = 100 μm. ( C ) Quantification of B. n = 3. ( D )The content of ADAM9 in the supernatant of GHRH neurons. Microglial supernatant was used as a positive control. n = 3. ( E ) CD11b and Iba-1 were coexpressed in the SAH group, and microglia were more strongly activated in the SAH group than in the sham‐operated group. CD11b is an M1 microglial marker. The arrow indicates microglia. Scale bar = 20 μm. ( F ) ADAM9 and Iba-1 were coexpressed in hypothalamic microglia in primary culture on day 7 after SAH surgery. Scale bar = 20 μm. ( G ) Caspase-1 was highly expressed in the ARC at 7 days after SAH surgery. GAPDH expression was used to confirm equal protein loading and blotting. ( H ) Quantification of G. n = 3. ( I ) Immunofluorescence showed that the number of GHRH + GSDMD + neurons in the SAH group was significantly greater than that in the control group. ( J ) Quantification of the ARC in I. n = 3. ( K ) The expression of IL-1β, IL-6 and IL-8 in the ARC on day 7 after SAH surgery was significantly higher than that in the sham‐operated mice. n = 3. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; *** p < 0.001

    Journal: Journal of Neuroinflammation

    Article Title: Microglia-derived ADAM9 promote GHRH neurons pyroptosis by Mad2L2-JNK-caspase-1 pathway in subarachnoid hemorrhage

    doi: 10.1186/s12974-024-03299-x

    Figure Lengend Snippet: ADAM9 induces pyroptosis in GHRH neurons in SAH. ( A ) The content of ADAM9 in the CSF of mice on day 7 after SAH surgery was significantly greater than that in the CSF of sham-operated mice. n = 3. ( B ) The expression of ADAM9 and the activation of microglia at 3 d, 7 d and 14 d after SAH surgery compared with those in sham‐operated mice. Iba-1 is a microglial marker. Scale bar = 100 μm. ( C ) Quantification of B. n = 3. ( D )The content of ADAM9 in the supernatant of GHRH neurons. Microglial supernatant was used as a positive control. n = 3. ( E ) CD11b and Iba-1 were coexpressed in the SAH group, and microglia were more strongly activated in the SAH group than in the sham‐operated group. CD11b is an M1 microglial marker. The arrow indicates microglia. Scale bar = 20 μm. ( F ) ADAM9 and Iba-1 were coexpressed in hypothalamic microglia in primary culture on day 7 after SAH surgery. Scale bar = 20 μm. ( G ) Caspase-1 was highly expressed in the ARC at 7 days after SAH surgery. GAPDH expression was used to confirm equal protein loading and blotting. ( H ) Quantification of G. n = 3. ( I ) Immunofluorescence showed that the number of GHRH + GSDMD + neurons in the SAH group was significantly greater than that in the control group. ( J ) Quantification of the ARC in I. n = 3. ( K ) The expression of IL-1β, IL-6 and IL-8 in the ARC on day 7 after SAH surgery was significantly higher than that in the sham‐operated mice. n = 3. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; *** p < 0.001

    Article Snippet: The following primary antibodies were used: anti-ADAM9 (Thermo Fisher Scientific, Cat# PA5-76732, RRID: AB_2720459), anti-ADAM9 (Santa Cruz, Cat# sc-135822, RRID: AB_2221762), anti-Iba1 (Wako, Cat# 011-27991, RRID: AB_2935833), anti-CD11b (Abcam, Cat# ab184308, RRID: AB_2889154), GHRH (Thermo Fisher Scientific, Cat# PA5-92408, RRID: AB_2806480), GSDMD (Proteintech, Cat# 66387-1-Ig, RRID: AB_2881763), MAD2L2 (BD Biosciences Cat# 612266, RRID: AB_399583)), GFAP (Novus Cat# NBP2-33184DL594, RRID: AB_2923514), Olig2 (Proteintech, Cat# 66513-1-Ig, RRID: AB_2881876), NeuN (Abcam, Cat# ab104224, RRID: AB_10711040), caspase-1 (Abcam, Cat# ab207802, RRID: AB_2889889), GAPDH (Abcam, Cat# ab8245, RRID: AB_2107448), P-JNK (CST, Cat# 9255 S, RRID: AB_2307321), JNK (CST, Cat# 9252 S, RRID: AB_2250373), P-ERK (CST, Cat# 4370 S, RRID: AB_2315112), ERK (CST, Cat# 4695 S, RRID: AB_390779), P-p38 (CST, Cat# 9216 S, RRID: AB_331296), and p38 (CST, Cat# 8690 S, RRID: AB_10999090).

    Techniques: Expressing, Activation Assay, Marker, Positive Control, Immunofluorescence, Control

    GH and cognitive changes in SAH mice after elimination of hypothalamic microglia. ( A ) Schematic representation of SAH surgery involving the elimination of microglia. ( B ) After the use of PLX3397, the number of microglia and the expression of ADAM9 decreased at 7 days after SAH surgery compared with those in control chow-fed mice. Scale bar = 100 μm. ( C ) Quantification of the data in B. n = 3. ( D ) After the use of PLX3397, the expression of GSDMD in GHRH neurons decreased on day 7 after SAH surgery. Scale bar = 100 μm. ( E ) Quantification of the ARC in D. n = 3. ( F ) After the use of PLX3397, the expression of caspase-1 in the ARC decreased at 7 days after SAH surgery. GAPDH expression was used to confirm equal protein loading and blotting. ( G ) Quantification of F. n = 3. ( H ) After treatment with PLX3397, the expression levels of 1β, IL-6 and IL-8 were decreased in the ARC at 7 days after SAH surgery. n = 3. ( I ) The expression of GH increased in the PLX3397 group. n = 3. ( J - M ) Cognitive ability improved in the PLX3397 group. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Journal: Journal of Neuroinflammation

    Article Title: Microglia-derived ADAM9 promote GHRH neurons pyroptosis by Mad2L2-JNK-caspase-1 pathway in subarachnoid hemorrhage

    doi: 10.1186/s12974-024-03299-x

    Figure Lengend Snippet: GH and cognitive changes in SAH mice after elimination of hypothalamic microglia. ( A ) Schematic representation of SAH surgery involving the elimination of microglia. ( B ) After the use of PLX3397, the number of microglia and the expression of ADAM9 decreased at 7 days after SAH surgery compared with those in control chow-fed mice. Scale bar = 100 μm. ( C ) Quantification of the data in B. n = 3. ( D ) After the use of PLX3397, the expression of GSDMD in GHRH neurons decreased on day 7 after SAH surgery. Scale bar = 100 μm. ( E ) Quantification of the ARC in D. n = 3. ( F ) After the use of PLX3397, the expression of caspase-1 in the ARC decreased at 7 days after SAH surgery. GAPDH expression was used to confirm equal protein loading and blotting. ( G ) Quantification of F. n = 3. ( H ) After treatment with PLX3397, the expression levels of 1β, IL-6 and IL-8 were decreased in the ARC at 7 days after SAH surgery. n = 3. ( I ) The expression of GH increased in the PLX3397 group. n = 3. ( J - M ) Cognitive ability improved in the PLX3397 group. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Article Snippet: The following primary antibodies were used: anti-ADAM9 (Thermo Fisher Scientific, Cat# PA5-76732, RRID: AB_2720459), anti-ADAM9 (Santa Cruz, Cat# sc-135822, RRID: AB_2221762), anti-Iba1 (Wako, Cat# 011-27991, RRID: AB_2935833), anti-CD11b (Abcam, Cat# ab184308, RRID: AB_2889154), GHRH (Thermo Fisher Scientific, Cat# PA5-92408, RRID: AB_2806480), GSDMD (Proteintech, Cat# 66387-1-Ig, RRID: AB_2881763), MAD2L2 (BD Biosciences Cat# 612266, RRID: AB_399583)), GFAP (Novus Cat# NBP2-33184DL594, RRID: AB_2923514), Olig2 (Proteintech, Cat# 66513-1-Ig, RRID: AB_2881876), NeuN (Abcam, Cat# ab104224, RRID: AB_10711040), caspase-1 (Abcam, Cat# ab207802, RRID: AB_2889889), GAPDH (Abcam, Cat# ab8245, RRID: AB_2107448), P-JNK (CST, Cat# 9255 S, RRID: AB_2307321), JNK (CST, Cat# 9252 S, RRID: AB_2250373), P-ERK (CST, Cat# 4370 S, RRID: AB_2315112), ERK (CST, Cat# 4695 S, RRID: AB_390779), P-p38 (CST, Cat# 9216 S, RRID: AB_331296), and p38 (CST, Cat# 8690 S, RRID: AB_10999090).

    Techniques: Expressing, Control

    ADAM9 induces pyroptosis in GHRH neurons via the JNK pathway. ( A ) Protein levels of JNK, ERK and p38 pathway markers were measured on day 7 postsurgery in SAH and sham-operated mice by Western blot. GAPDH was used as a loading control. ( B-G ) Quantification of A. n = 3. ( H ) The expression of GSDMD in GHRH neurons was increased when ADAM9 was injected into the ARC. Scale bar = 100 μm. ( I ) Quantification of the ARC in H. n = 3. ( J ) Schematic representation of SAH surgery after blocking the JNK pathway. ( K ) After SP600125 treatment, the expression of GSDMD in GHRH neurons decreased at 7 days after SAH surgery. Scale bar = 100 μm. ( L ) Quantification of the ARC in K. n = 3. ( M ) After the use of SP600125, the protein levels of JNK and pyroptosis pathway markers were measured on day 7 postsurgery in SAH and sham‐operated mice by Western blotting. GAPDH was used as a loading control. ( N - P ) Quantification of M. n = 3. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; *** p < 0.001

    Journal: Journal of Neuroinflammation

    Article Title: Microglia-derived ADAM9 promote GHRH neurons pyroptosis by Mad2L2-JNK-caspase-1 pathway in subarachnoid hemorrhage

    doi: 10.1186/s12974-024-03299-x

    Figure Lengend Snippet: ADAM9 induces pyroptosis in GHRH neurons via the JNK pathway. ( A ) Protein levels of JNK, ERK and p38 pathway markers were measured on day 7 postsurgery in SAH and sham-operated mice by Western blot. GAPDH was used as a loading control. ( B-G ) Quantification of A. n = 3. ( H ) The expression of GSDMD in GHRH neurons was increased when ADAM9 was injected into the ARC. Scale bar = 100 μm. ( I ) Quantification of the ARC in H. n = 3. ( J ) Schematic representation of SAH surgery after blocking the JNK pathway. ( K ) After SP600125 treatment, the expression of GSDMD in GHRH neurons decreased at 7 days after SAH surgery. Scale bar = 100 μm. ( L ) Quantification of the ARC in K. n = 3. ( M ) After the use of SP600125, the protein levels of JNK and pyroptosis pathway markers were measured on day 7 postsurgery in SAH and sham‐operated mice by Western blotting. GAPDH was used as a loading control. ( N - P ) Quantification of M. n = 3. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; *** p < 0.001

    Article Snippet: The following primary antibodies were used: anti-ADAM9 (Thermo Fisher Scientific, Cat# PA5-76732, RRID: AB_2720459), anti-ADAM9 (Santa Cruz, Cat# sc-135822, RRID: AB_2221762), anti-Iba1 (Wako, Cat# 011-27991, RRID: AB_2935833), anti-CD11b (Abcam, Cat# ab184308, RRID: AB_2889154), GHRH (Thermo Fisher Scientific, Cat# PA5-92408, RRID: AB_2806480), GSDMD (Proteintech, Cat# 66387-1-Ig, RRID: AB_2881763), MAD2L2 (BD Biosciences Cat# 612266, RRID: AB_399583)), GFAP (Novus Cat# NBP2-33184DL594, RRID: AB_2923514), Olig2 (Proteintech, Cat# 66513-1-Ig, RRID: AB_2881876), NeuN (Abcam, Cat# ab104224, RRID: AB_10711040), caspase-1 (Abcam, Cat# ab207802, RRID: AB_2889889), GAPDH (Abcam, Cat# ab8245, RRID: AB_2107448), P-JNK (CST, Cat# 9255 S, RRID: AB_2307321), JNK (CST, Cat# 9252 S, RRID: AB_2250373), P-ERK (CST, Cat# 4370 S, RRID: AB_2315112), ERK (CST, Cat# 4695 S, RRID: AB_390779), P-p38 (CST, Cat# 9216 S, RRID: AB_331296), and p38 (CST, Cat# 8690 S, RRID: AB_10999090).

    Techniques: Western Blot, Control, Expressing, Injection, Blocking Assay

    Changes in GHRH neurons after MAD2L2 KO in SAH mice. ( A ) The expression of MAD2L2 in GHRH neurons was higher on day 7 postsurgery in SAH mice than in sham-operated mice. Scale bar = 100 μm. ( B ) Quantification of the ARC in A. n = 3. ( C ) The expression of MAD2L2 in GHRH neurons was higher when ADAM9 was injected into the ARC than when it was injected into the PBS. Scale bar = 100 μm. ( D ) Quantification of the ARC in C. n = 3. ( E ) After MAD2L2 KO, the protein levels of the JNK pathway markers caspase-1 and MAD2L2 were measured on day 7 postsurgery in SAH mice by Western blotting. GAPDH was used as a loading control. ( F - I ) Quantification of E. n = 3. ( J ) After MAD2L2 KO, the expression of MAD2L2 in GHRH neurons was decreased on day 7 postsurgery in SAH mice. Scale bar = 100 μm. ( K ) Quantification of the ARC in J. n = 3. ( L ) After MAD2L2 KO, the expression of GSDMD in GHRH neurons was decreased on day 7 postsurgery in SAH mice. Scale bar = 100 μm. ( M ) Quantification of the ARC in L. n = 3. ( N )The expression of GH increased in the KO MAD2L2 + SAH group. n = 3. ( O-R ) Cognitive ability improved in the KO MAD2L2 + SAH group. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Journal: Journal of Neuroinflammation

    Article Title: Microglia-derived ADAM9 promote GHRH neurons pyroptosis by Mad2L2-JNK-caspase-1 pathway in subarachnoid hemorrhage

    doi: 10.1186/s12974-024-03299-x

    Figure Lengend Snippet: Changes in GHRH neurons after MAD2L2 KO in SAH mice. ( A ) The expression of MAD2L2 in GHRH neurons was higher on day 7 postsurgery in SAH mice than in sham-operated mice. Scale bar = 100 μm. ( B ) Quantification of the ARC in A. n = 3. ( C ) The expression of MAD2L2 in GHRH neurons was higher when ADAM9 was injected into the ARC than when it was injected into the PBS. Scale bar = 100 μm. ( D ) Quantification of the ARC in C. n = 3. ( E ) After MAD2L2 KO, the protein levels of the JNK pathway markers caspase-1 and MAD2L2 were measured on day 7 postsurgery in SAH mice by Western blotting. GAPDH was used as a loading control. ( F - I ) Quantification of E. n = 3. ( J ) After MAD2L2 KO, the expression of MAD2L2 in GHRH neurons was decreased on day 7 postsurgery in SAH mice. Scale bar = 100 μm. ( K ) Quantification of the ARC in J. n = 3. ( L ) After MAD2L2 KO, the expression of GSDMD in GHRH neurons was decreased on day 7 postsurgery in SAH mice. Scale bar = 100 μm. ( M ) Quantification of the ARC in L. n = 3. ( N )The expression of GH increased in the KO MAD2L2 + SAH group. n = 3. ( O-R ) Cognitive ability improved in the KO MAD2L2 + SAH group. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Article Snippet: The following primary antibodies were used: anti-ADAM9 (Thermo Fisher Scientific, Cat# PA5-76732, RRID: AB_2720459), anti-ADAM9 (Santa Cruz, Cat# sc-135822, RRID: AB_2221762), anti-Iba1 (Wako, Cat# 011-27991, RRID: AB_2935833), anti-CD11b (Abcam, Cat# ab184308, RRID: AB_2889154), GHRH (Thermo Fisher Scientific, Cat# PA5-92408, RRID: AB_2806480), GSDMD (Proteintech, Cat# 66387-1-Ig, RRID: AB_2881763), MAD2L2 (BD Biosciences Cat# 612266, RRID: AB_399583)), GFAP (Novus Cat# NBP2-33184DL594, RRID: AB_2923514), Olig2 (Proteintech, Cat# 66513-1-Ig, RRID: AB_2881876), NeuN (Abcam, Cat# ab104224, RRID: AB_10711040), caspase-1 (Abcam, Cat# ab207802, RRID: AB_2889889), GAPDH (Abcam, Cat# ab8245, RRID: AB_2107448), P-JNK (CST, Cat# 9255 S, RRID: AB_2307321), JNK (CST, Cat# 9252 S, RRID: AB_2250373), P-ERK (CST, Cat# 4370 S, RRID: AB_2315112), ERK (CST, Cat# 4695 S, RRID: AB_390779), P-p38 (CST, Cat# 9216 S, RRID: AB_331296), and p38 (CST, Cat# 8690 S, RRID: AB_10999090).

    Techniques: Expressing, Injection, Western Blot, Control

    The effect of sorafenib on SAH mice. ( A ) There was no significant change in the number of microglia in SAH mice after the administration of sorafenib. However, the expression of ADAM9 decreased. Scale bar = 100 μm. ( B ) Quantification of the ARC in A. n = 3. ( C ) The expression of MAD2L2 in GHRH neurons was decreased in SAH mice after treatment with sorafenib. Scale bar = 100 μm. ( D ) Quantification of the ARC in C. n = 3. ( E ) The expression of GSDMD in GHRH neurons was decreased in SAH mice after the administration of sorafenib. Scale bar = 100 μm. ( F ) Quantification of the ARC in E. n = 3. ( G ) After treatment with sorafenib, the protein levels of JNK pathway markers and caspase-1 were measured in SAH mice by Western blotting. GAPDH was used as a loading control. ( H-J ) Quantification of G. n = 3. ( K ) After treatment with sorafenib, the expression levels of 1β, IL-6 and IL-8 were decreased in the ARC of SAH mice. n = 3. ( L ) After the use of sorafenib, the expression of GH increased in SAH mice. n = 3. ( M-P ) After the use of sorafenib, the cognitive ability of SAH mice improved. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Journal: Journal of Neuroinflammation

    Article Title: Microglia-derived ADAM9 promote GHRH neurons pyroptosis by Mad2L2-JNK-caspase-1 pathway in subarachnoid hemorrhage

    doi: 10.1186/s12974-024-03299-x

    Figure Lengend Snippet: The effect of sorafenib on SAH mice. ( A ) There was no significant change in the number of microglia in SAH mice after the administration of sorafenib. However, the expression of ADAM9 decreased. Scale bar = 100 μm. ( B ) Quantification of the ARC in A. n = 3. ( C ) The expression of MAD2L2 in GHRH neurons was decreased in SAH mice after treatment with sorafenib. Scale bar = 100 μm. ( D ) Quantification of the ARC in C. n = 3. ( E ) The expression of GSDMD in GHRH neurons was decreased in SAH mice after the administration of sorafenib. Scale bar = 100 μm. ( F ) Quantification of the ARC in E. n = 3. ( G ) After treatment with sorafenib, the protein levels of JNK pathway markers and caspase-1 were measured in SAH mice by Western blotting. GAPDH was used as a loading control. ( H-J ) Quantification of G. n = 3. ( K ) After treatment with sorafenib, the expression levels of 1β, IL-6 and IL-8 were decreased in the ARC of SAH mice. n = 3. ( L ) After the use of sorafenib, the expression of GH increased in SAH mice. n = 3. ( M-P ) After the use of sorafenib, the cognitive ability of SAH mice improved. n = 10. ARC: Arcuate nucleus. 3 V: The third ventricle. * p < 0.05; ** p < 0.01; *** p < 0.001

    Article Snippet: The following primary antibodies were used: anti-ADAM9 (Thermo Fisher Scientific, Cat# PA5-76732, RRID: AB_2720459), anti-ADAM9 (Santa Cruz, Cat# sc-135822, RRID: AB_2221762), anti-Iba1 (Wako, Cat# 011-27991, RRID: AB_2935833), anti-CD11b (Abcam, Cat# ab184308, RRID: AB_2889154), GHRH (Thermo Fisher Scientific, Cat# PA5-92408, RRID: AB_2806480), GSDMD (Proteintech, Cat# 66387-1-Ig, RRID: AB_2881763), MAD2L2 (BD Biosciences Cat# 612266, RRID: AB_399583)), GFAP (Novus Cat# NBP2-33184DL594, RRID: AB_2923514), Olig2 (Proteintech, Cat# 66513-1-Ig, RRID: AB_2881876), NeuN (Abcam, Cat# ab104224, RRID: AB_10711040), caspase-1 (Abcam, Cat# ab207802, RRID: AB_2889889), GAPDH (Abcam, Cat# ab8245, RRID: AB_2107448), P-JNK (CST, Cat# 9255 S, RRID: AB_2307321), JNK (CST, Cat# 9252 S, RRID: AB_2250373), P-ERK (CST, Cat# 4370 S, RRID: AB_2315112), ERK (CST, Cat# 4695 S, RRID: AB_390779), P-p38 (CST, Cat# 9216 S, RRID: AB_331296), and p38 (CST, Cat# 8690 S, RRID: AB_10999090).

    Techniques: Expressing, Western Blot, Control

    Journal: iScience

    Article Title: Distinct effects of sacituzumab govitecan and berzosertib on DNA damage response in ovarian cancer

    doi: 10.1016/j.isci.2024.111283

    Figure Lengend Snippet:

    Article Snippet: Rabbit anti- MAD2L2/REV7 , ProteinTech , Cat #12683-1-AP; RRID: AB_2139530.

    Techniques: Recombinant, Staining, Western Blot, Stripping, Control, SYBR Green Assay, cDNA Synthesis, Gene Expression, Software, Microscopy, Imaging