Journal: Cancer letters

Article Title: Anti-tumor activity of CDYL2b in prostate cancer

doi: 10.1016/j.canlet.2025.217987

Figure Lengend Snippet: Inhibition of CDYL2b expression by JMJD2A and JMJD2B. ( A ) Downregulation of JMJD2A with two different shRNAs in DU145 prostate cancer cells led to enhanced CDYL2b mRNA levels; one-way ANOVA with Tukey’s multiple comparisons test (n = 3; ****, P < 0.0001). Bottom two panels show Western blots for JMJD2A and the loading control GAPDH. (B) Analogous upon JMJD2B downregulation; one-way ANOVA with Tukey’s multiple comparisons test (n = 3; **. P < 0.01; ***, P < 0.001). ( C ) Exon-intron structure of the four predicted transcripts derived from the human CDYL2 gene on chromosome 16 (left). All transcripts have a unique first exon, but share the subsequent six exons. The locations of the start ( ATG ) and stop ( TGA ) codons are indicated and the corresponding four CDYL2 protein isoforms are depicted on the right. CDYL2a-c share 498 C-terminal amino acids, while CDYL2d shares all of its 447 amino acids with the other three isoforms. CD, chromodomain; CECH, crotonase/enoyl-coenzyme A hydratase domain. ( D ) RT-PCR for the four different CDYL2 transcript variants in DU145 cells. For CDYL2a and CDYL2c , no Ct values were observable at the experimentally reliable maximum of 39 PCR cycles; unpaired, two-tailed t -test (n = 3; ****, P < 0.0001). ( E ) Chromatin immunoprecipitation assay with DU145 cells. The scheme at the top shows the respective first exons of CDYL2a and CDYL2b , with +1 designating the transcription start site of the latter and the location of the amplified upstream and downstream genomic DNA fragments being indicated by brown, horizontal bars.

Article Snippet: Chromatin was sheared through sonication, debris removed by centrifugation, immunoprecipitations performed with no antibody, IgG control (sc-2025; Santa Cruz Biotechnology, Santa Cruz, CA, USA), JMJD2A antibodies (09–809; Millipore, Temecula, CA, USA) or JMJD2B antibodies (A301–477A; Bethyl Laboratories, Montgomery, TX, USA) and bound DNA recovered after reverse-crosslinking [ ].

Techniques: Inhibition, Expressing, Western Blot, Control, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test, Chromatin Immunoprecipitation, Amplification

Journal: Cancer letters

Article Title: Anti-tumor activity of CDYL2b in prostate cancer

doi: 10.1016/j.canlet.2025.217987

Figure Lengend Snippet: Interaction of CDYL2b with JMJD2B. ( A ) Indicated 6Myc-tagged JMJD proteins were expressed without and with Flag-CDYL2b. Immunoprecipitation (IP) with anti-Flag antibodies was followed by anti -Myc Western blotting to reveal coprecipitated JMJD proteins (top). Middle and bottom panels show input levels of 6Myc-tagged JMJD proteins and of Flag-tagged CDYL2b, respectively. ( B ) Gene activity assays with the TORU luciferase reporter construct in DU145 prostate cancer cells. Expression of ETV1, JMJD2B and/or CDYL2b is indicated. ( C ) Likewise, with a YAP1 luciferase reporter construct. Statistical significance was assessed with one-way ANOVA (Tukey’s multiple comparisons test; n = 4); ns, not significant; *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Article Snippet: Chromatin was sheared through sonication, debris removed by centrifugation, immunoprecipitations performed with no antibody, IgG control (sc-2025; Santa Cruz Biotechnology, Santa Cruz, CA, USA), JMJD2A antibodies (09–809; Millipore, Temecula, CA, USA) or JMJD2B antibodies (A301–477A; Bethyl Laboratories, Montgomery, TX, USA) and bound DNA recovered after reverse-crosslinking [ ].

Techniques: Immunoprecipitation, Western Blot, Activity Assay, Luciferase, Construct, Expressing

Journal: Cancer letters

Article Title: Anti-tumor activity of CDYL2b in prostate cancer

doi: 10.1016/j.canlet.2025.217987

Figure Lengend Snippet: Model. The tumor suppressive action of CDYL2b entails the upregulation of HES7 , KLF17 and TBX6 ; please note that HES7 may (also) be activated by TBX6, thereby establishing an indirect way of how CDYL2b could stimulate HES7 transcription. Suppression of CDYL2b transcription by JMJD2A and JMJD2B and/or inhibition of CDYL2b-mediated HES7 transcription through JMJD2B, which may involve complex formation between JMJD2B and CDYL2b, constrains the anti-oncogenic functions of CDYL2b.

Article Snippet: Chromatin was sheared through sonication, debris removed by centrifugation, immunoprecipitations performed with no antibody, IgG control (sc-2025; Santa Cruz Biotechnology, Santa Cruz, CA, USA), JMJD2A antibodies (09–809; Millipore, Temecula, CA, USA) or JMJD2B antibodies (A301–477A; Bethyl Laboratories, Montgomery, TX, USA) and bound DNA recovered after reverse-crosslinking [ ].

Techniques: Inhibition