cleaved parp  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Electrostatic control of chromatin compaction safeguards against apoptotic DNA release

doi: 10.64898/2026.02.23.707452

Figure Lengend Snippet: a-d , Fluorescence-based western blot of histone acetylation levels of four core histones during apoptosis and mitosis. Cell lysates were harvested from synchronous interphase, apoptotic and mitotic cultures, and levels of histone acetylation were measured relative to core histones. Cleaved PARP and MPM2 were blotted to detect apoptotic and mitotic states, respectively. e-h , Quantification of histone acetylation levels, relative to core histone amounts in the same loading lane. n =5 for all conditions. Bars indicate mean; significance was tested by a parametric ratio paired t-test (H2A K5-Ac, P =0.0016 (apoptosis) P =0.0007 (mitosis); H2B K12-Ac, P =0.0122 (apoptosis) P =0.0692 (mitosis); H3 K27-Ac, P =0.0294 (apoptosis) P =0.0218 (mitosis); H4 K16-Ac, P =0.0036 (apoptosis) P =0.0498 (mitosis). i, j , Imaging of chromatin density in the presence and absence of active histone deacetylases in interphase, apoptosis and mitosis. Chromatin was stained with the DNA dye SPY650, and Annexin V-488 was used to detect loss of membrane asymmetry in apoptosis. Scale bar 10 µm. k, l , Quantification of chromatin compaction. n =131 for interphase, n =124 for apoptosis, n =84 for mitosis (control); n =105 for interphase, n =121 for apoptosis, n =95 for mitosis (romidepsin). Bars indicate mean; significance was tested by a two-tailed Mann-Whitney test (control apoptosis vs interphase, P =<0.0001; control mitosis vs interphase, P =<0.0001; control apoptosis vs mitosis, P =<0.0001; romidepsin-treated apoptosis vs control apoptosis, P =<0.0001; romidepsin-treated mitosis vs control mitosis, P =<0.0001; romidepsin-treated apoptosis vs romidepsin-treated mitosis, P =<0.0001).

Article Snippet: For immunofluorescence, wells were blocked with 5% BSA in PBS for 30 min with rocking before primary antibody Cleaved PARP (Asp214) (D64E10) XP® Rabbit (Cell Signaling Technologies; #5625S; Lot 18) was added at 1:400 dilution for 2 h with rocking.

Techniques: Fluorescence, Western Blot, Imaging, Staining, Membrane, Control, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Electrostatic control of chromatin compaction safeguards against apoptotic DNA release

doi: 10.64898/2026.02.23.707452

Figure Lengend Snippet: a-d , Fluorescence-based western blot of histone acetylation levels of four core histones during apoptosis and mitosis, in the presence and absence of romidepsin. Cell lysates were harvested from control and romidepsin-treated interphase, apoptotic and mitotic cultures, and levels of histone acetylation were measured relative to core histones. Cleaved PARP and MPM2 were blotted to detect apoptotic and mitotic states, respectively, in both control and romidepsin-treated conditions. e-h , Quantification of histone acetylation levels, relative to core histone amounts in the same loading lane. n =4 for all conditions. Bars indicate mean; significance was tested by a parametric ratio paired t-test (H2A K5-Ac, P =0.0412 (interphase), P =0.0047 (apoptosis), P =0.0004 (mitosis); H2B K12-Ac, P =0.0003 (interphase), P =0.0005 (apoptosis), P =0.0009 (mitosis); H3 K27-Ac, P =<0.0001 (interphase), P =0.0028 (apoptosis), P =0.0004 (mitosis); H4 K16-Ac, P =0.0333 (interphase), P =0.0037 (apoptosis), P =0.0166 (mitosis)).

Article Snippet: For immunofluorescence, wells were blocked with 5% BSA in PBS for 30 min with rocking before primary antibody Cleaved PARP (Asp214) (D64E10) XP® Rabbit (Cell Signaling Technologies; #5625S; Lot 18) was added at 1:400 dilution for 2 h with rocking.

Techniques: Fluorescence, Western Blot, Control

Journal: bioRxiv

Article Title: Electrostatic control of chromatin compaction safeguards against apoptotic DNA release

doi: 10.64898/2026.02.23.707452

Figure Lengend Snippet: a , Agarose gel electrophoresis after PCR amplification of the CAD genomic locus in wild-type and CAD KO HeLa cells. * indicates expected length of wild-type locus, ** indicates expected length of locus after CRISPR/Cas9-mediated partial deletion. b , Agarose gel electrophoresis of fragmented DNA isolated from wild-type and CAD KO cells in the presence and absence of romidepsin and BH3 mimetics. c , Imaging of fixed wild-type and CAD KO cells, stained for cleaved PARP and TUNEL signal. DNA was stained with SPY650. Scale bar 5 µm. d , Quantification of TUNEL signal normalised to chromatin density ( d ) and chromatin density ( e ) in fixed live and apoptotic wild-type and CAD KO cells, in the presence and absence of romidepsin. n =76 for live wild-type, n =103 for apoptotic wild-type, n =103 for apoptotic CAD KO (control); n =63 for live wild-type, n =77 for apoptotic wild-type, n =103 for apoptotic CAD KO (romidepsin). Bars indicate mean; significance was tested by a two-tailed Mann-Whitney test (apoptotic wild-type vs live wild-type, P =<0.0001 (TUNEL), P =<0.0001 (chromatin); apoptotic CAD KO vs apoptotic wild-type, P =<0.0001 (TUNEL), romidepsin-treated apoptotic wild-type vs control apoptotic wild-type, P =<0.0001 (chromatin)).

Article Snippet: For immunofluorescence, wells were blocked with 5% BSA in PBS for 30 min with rocking before primary antibody Cleaved PARP (Asp214) (D64E10) XP® Rabbit (Cell Signaling Technologies; #5625S; Lot 18) was added at 1:400 dilution for 2 h with rocking.

Techniques: Agarose Gel Electrophoresis, Amplification, CRISPR, Isolation, Imaging, Staining, TUNEL Assay, Control, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Electrostatic control of chromatin compaction safeguards against apoptotic DNA release

doi: 10.64898/2026.02.23.707452

Figure Lengend Snippet: a-d , Fluorescence-based western blot of histone acetylation levels of four core histones during apoptosis in Jurkat cells, in the presence and absence of romidepsin. Jurkat cell lysates were harvested from control and romidepsin-treated interphase and apoptotic cultures, and levels of histone acetylation were measured relative to core histones. Cleaved PARP was blotted to detect an apoptotic state in both control and romidepsin-treated conditions. e-h , Quantification of Jurkat cell histone acetylation levels, relative to core histone amounts in the same loading lane. n =5 for all conditions. Bars indicate mean; significance was tested by a parametric ratio paired t-test (H2A K5-Ac, P =0.0215 (control apoptosis vs control interphase), P =0.0032 (romidepsin-treated apoptosis vs control apoptosis); H2B K12-Ac, P =0.0032 (control apoptosis vs control interphase), P =<0.0001 (romidepsin-treated apoptosis vs control apoptosis); H3 K27-Ac, P =0.0235 (control apoptosis vs control interphase), P =0.0002 (romidepsin-treated apoptosis vs control apoptosis); H4 K16-Ac, P =0.0452 (control apoptosis vs control interphase), P =0.0069 (romidepsin-treated apoptosis vs control apoptosis)). i , Imaging of chromatin density in apoptotic wild-type and CAD KO Jurkat cells in the presence and absence of active histone deacetylases. Chromatin was stained with the DNA dye SPY650, and Annexin V-488 was used to detect loss of membrane asymmetry in apoptosis. Contours were drawn from the outer contour of Annexin V staining. Scale bar 10 µm. b , Quantification of chromatin compaction in apoptotic Jurkat cells. n =94 for wild-type control apoptosis, n =93 for wild-type romidepsin-treated apoptosis, n =86 for CAD KO control apoptosis, n =73 for CAD KO romidepsin-treated apoptosis. Bars indicate mean; significance was tested by a two-tailed Mann-Whitney test (romidepsin-treated wild-type apoptosis vs control wild-type apoptosis, P =<0.0001; romidepsin-treated CAD KO apoptosis vs control CAD KO apoptosis, P =<0.0001; romidepsin-treated CAD KO apoptosis vs romidepsin-treated wild-type apoptosis, P =<0.0001). k , Flow cytometry plots of DNA content in ApoEVs from apoptotic wild-type and CAD KO Jurkat cells, in the presence and absence of romidepsin. ApoEVs were identified based on size and Annexin V-Pacific Blue staining, DNA was stained with SPY650. Positive populations were defined by unstained controls. DNA percentages displayed as mean values across three biological replicates. l , Quantification of Jurkat ApoEV DNA content, as a percentage of all ApoEVs. n =3 for all conditions. Bars indicate mean; significance was tested by a parametric paired t-test (DNA content in romidepsin-treated wild-type cells, P =0.0186).

Article Snippet: For immunofluorescence, wells were blocked with 5% BSA in PBS for 30 min with rocking before primary antibody Cleaved PARP (Asp214) (D64E10) XP® Rabbit (Cell Signaling Technologies; #5625S; Lot 18) was added at 1:400 dilution for 2 h with rocking.

Techniques: Fluorescence, Western Blot, Control, Imaging, Staining, Membrane, Two Tailed Test, MANN-WHITNEY, Flow Cytometry