rabbit anti erm  (Cell Signaling Technology Inc)

Journal: Oncogene

Article Title: The CIC::DUX4 oncoprotein maintains DNA integrity through direct regulation of the catalytic subunit of DNA polymerase epsilon (POLE)

doi: 10.1038/s41388-025-03507-9

Figure Lengend Snippet: A ChIP-seq track from NCC_CDS1_X1_C1 demonstrating CIC::DUX4 occupancy (upper gray and blue) on intron 1 of POLE . The identified peak within the intronic region of POLE on chromosome 12 containing the canonical CIC::DNA binding motif TGAATGAA and multiple variant TGAATGAG motifs. Immunoglobulin (IgG) served as control. B Schematic of the luciferase reporter construct, PGL4.10-POLE that contains the genomic sequence of POLE that is occupied by CIC::DUX4. C Relative luciferase activity in HEK293T expressing PGL4.10, PGL4.10-POLE, PGL4.10 + CIC::DUX4, and PGL4.10-POLE + CIC::DUX4 constructs. D Relative mRNA expression levels of CIC , POLE , and ETV4 in NCC_CDS1_X1_C1 (top panels) and NCC_CDS2_C1 (bottom panels) cells transfected with siCIC (CIC::DUX4 knockdown) or scramble control (siCON). E Immunoblots of CIC, POLE and ETV4 from NCC_CDS1_X1_C1 and NCC_CDS2_C1 cells transfected with siCIC or siCON for 48 h. F Relative mRNA expression levels of CIC, POLE , and ETV4 in NCC_CDS1_X1_C1 (left panel) and NCC_CDS2_C1 (right panel) cells over-expressing CIC::DUX4-HA or empty vector (EV) control. Error bars represent standard deviation (SD); statistical significance was assessed Student’s t test. *P < 0.05 , **P < 0.01 , ***P < 0.001, ****P < 0.0001. Data represent results from three independent experiments.

Article Snippet: The membranes were blocked in 5% BSA in TBS-T and then probed with primary antibodies against POLE (Santa Cruz Biotechnology, sc-390785, 1:1000), p21 Waf1/Cip1 (12D1) (Cell Signaling Technology (CST) 2947, 1:500), p53 (CST, 9282, 1:1000), Anti-Pea3 (ETV4) (San Cruz, PEA3 Antibody [ ]: sc-113, 1:500), ETV5 (CST, 16274, 1:1000), DUX4 clone P4H2 (Invitrogen MA5-16147, 1:1000), Phosph-Histon H2AX (Ser139) (CST, 2577, 1;500), Capicua CIC (Abcam, ab123822, 1:500), ACTIN (CST, 4970, 1:1000), Phospho-CHK1 ser345 (133D3) (CST,2348, 1:1000) and CHK1 (CST, 2360, 1:1000), HA-tag C29F4 (CST, 3724, 1:1000).

Techniques: ChIP-sequencing, Binding Assay, Variant Assay, Control, Luciferase, Construct, Sequencing, Activity Assay, Expressing, Transfection, Knockdown, Western Blot, Plasmid Preparation, Standard Deviation