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ZCWPW2 promotes lactylation and stability of recombination-associated proteins through coordination with ZCWPW1 in cultured cells. ( A ) Co-IP analysis of HEK293T cells overexpressed with HA-recombination-associated proteins (HSPA2, SYCP1, and MDC1) and Flag-ZCWPW2, with or without Myc-ZCWPW1, and in the presence or absence of siRNA targeting ZCWPW1 . Bar graph quantifies Kla signal intensity for each condition ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( B ) Co-IP analysis (left) for Kla of endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells after individual or combined knockdown of Zcwpw1 or/and Zcwpw2 using siRNAs. Quantification of Kla levels (right) showed a significant decrease in Kla upon Zcwpw1 or Zcwpw2 knockdown, with the greatest reduction observed upon dual silencing ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( C and D ) Co-IP (left) and quantification analysis (right) of Kla on endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells treated with the EP300 inhibitor <t>A-485,</t> the LDHA inhibitor sodium oxamate, or both (C), and with EP300 activator CTPB or exogenous lactate (D) ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.).
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ZCWPW2 promotes lactylation and stability of recombination-associated proteins through coordination with ZCWPW1 in cultured cells. ( A ) Co-IP analysis of HEK293T cells overexpressed with HA-recombination-associated proteins (HSPA2, SYCP1, and MDC1) and Flag-ZCWPW2, with or without Myc-ZCWPW1, and in the presence or absence of siRNA targeting ZCWPW1 . Bar graph quantifies Kla signal intensity for each condition ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( B ) Co-IP analysis (left) for Kla of endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells after individual or combined knockdown of Zcwpw1 or/and Zcwpw2 using siRNAs. Quantification of Kla levels (right) showed a significant decrease in Kla upon Zcwpw1 or Zcwpw2 knockdown, with the greatest reduction observed upon dual silencing ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( C and D ) Co-IP (left) and quantification analysis (right) of Kla on endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells treated with the EP300 inhibitor <t>A-485,</t> the LDHA inhibitor sodium oxamate, or both (C), and with EP300 activator CTPB or exogenous lactate (D) ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.).
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ZCWPW2 promotes lactylation and stability of recombination-associated proteins through coordination with ZCWPW1 in cultured cells. ( A ) Co-IP analysis of HEK293T cells overexpressed with HA-recombination-associated proteins (HSPA2, SYCP1, and MDC1) and Flag-ZCWPW2, with or without Myc-ZCWPW1, and in the presence or absence of siRNA targeting ZCWPW1 . Bar graph quantifies Kla signal intensity for each condition ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( B ) Co-IP analysis (left) for Kla of endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells after individual or combined knockdown of Zcwpw1 or/and Zcwpw2 using siRNAs. Quantification of Kla levels (right) showed a significant decrease in Kla upon Zcwpw1 or Zcwpw2 knockdown, with the greatest reduction observed upon dual silencing ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( C and D ) Co-IP (left) and quantification analysis (right) of Kla on endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells treated with the EP300 inhibitor <t>A-485,</t> the LDHA inhibitor sodium oxamate, or both (C), and with EP300 activator CTPB or exogenous lactate (D) ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.).
A 485 Medchemexpress, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ZCWPW2 promotes lactylation and stability of recombination-associated proteins through coordination with ZCWPW1 in cultured cells. ( A ) Co-IP analysis of HEK293T cells overexpressed with HA-recombination-associated proteins (HSPA2, SYCP1, and MDC1) and Flag-ZCWPW2, with or without Myc-ZCWPW1, and in the presence or absence of siRNA targeting ZCWPW1 . Bar graph quantifies Kla signal intensity for each condition ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( B ) Co-IP analysis (left) for Kla of endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells after individual or combined knockdown of Zcwpw1 or/and Zcwpw2 using siRNAs. Quantification of Kla levels (right) showed a significant decrease in Kla upon Zcwpw1 or Zcwpw2 knockdown, with the greatest reduction observed upon dual silencing ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( C and D ) Co-IP (left) and quantification analysis (right) of Kla on endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells treated with the EP300 inhibitor A-485, the LDHA inhibitor sodium oxamate, or both (C), and with EP300 activator CTPB or exogenous lactate (D) ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.).

Journal: Nucleic Acids Research

Article Title: A novel dual histone mark reader ZCWPW2 regulates meiotic recombination through lactylation and transcriptional regulation in humans and mice

doi: 10.1093/nar/gkag049

Figure Lengend Snippet: ZCWPW2 promotes lactylation and stability of recombination-associated proteins through coordination with ZCWPW1 in cultured cells. ( A ) Co-IP analysis of HEK293T cells overexpressed with HA-recombination-associated proteins (HSPA2, SYCP1, and MDC1) and Flag-ZCWPW2, with or without Myc-ZCWPW1, and in the presence or absence of siRNA targeting ZCWPW1 . Bar graph quantifies Kla signal intensity for each condition ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( B ) Co-IP analysis (left) for Kla of endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells after individual or combined knockdown of Zcwpw1 or/and Zcwpw2 using siRNAs. Quantification of Kla levels (right) showed a significant decrease in Kla upon Zcwpw1 or Zcwpw2 knockdown, with the greatest reduction observed upon dual silencing ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.). ( C and D ) Co-IP (left) and quantification analysis (right) of Kla on endogenous Mdc1, Hspa2, and Sycp1 in GC-2 cells treated with the EP300 inhibitor A-485, the LDHA inhibitor sodium oxamate, or both (C), and with EP300 activator CTPB or exogenous lactate (D) ( n = 3 independent experiments; two-tailed Student’s t -test; * P < 0.05; error bars, s.e.m.).

Article Snippet: HEK293T and GC-2 cells were treated either with 20 mM sodium oxamate (Santa Cruz, sc-215880), a specific LDHA inhibitor [ ], or with 10 μM A-485 (MedChemEpress, HY-107455), an EP300 inhibitor, for 16 h [ ].

Techniques: Cell Culture, Co-Immunoprecipitation Assay, Two Tailed Test, Knockdown