rabbit anti dnmt1  (Boster Bio)

Journal: NAR Cancer

Article Title: GSK-3484862, a DNMT1 degrader, promotes DNMT3B expression in lung cancer cells

doi: 10.1093/narcan/zcaf018

Figure Lengend Snippet: GSK-3484862 (labeled as GSK862) depletes DNMT1 protein and increases DNMT3B protein. ( A ) Mass spectrometry analysis of total 5mC content in A549 cells expressed as a percentage (i.e. number of 5mC-modified residues divided by the total number of cytosine residues × 100). The DMSO controls at different time points were averaged. ( B ) GSK862 induced hypomethylation, as determined by pyrosequencing, at three genomic loci ( CDX1 , NKX2-6 , and RASSF1A ) and at multiple transposable LINE-1 elements in A549 cells, as a function of treatment time and variation of compound concentrations. ( C ) Western blots showing endogenous levels of DNMT1 and DNMT3B in A549 cells following a 2-day treatment with GSK862 concentrations ranging from 0.02 to 2.0 μM. Note that the sample amount in the first lane was half of that in the other lanes. ( D ) Western blots showing increases of DNMT3B isoforms (indicated by arrows for long and short) and no changes in DNMT3A after treatment with GSK862. ( E ) Plots of intensities of protein bands of DNMT3A and DNMT3B shown in panel (D). ( F ) Concentration-dependent effect of GSK-3685032 (labeled as GSK032) in A549 cells. ( G ) Colony formation assay of A549 cells treated at the indicated doses of GSK862 for 10 days.

Article Snippet: The following primary antibodies were used in this study: DNMT1 [Cell Signaling Technology (CST), Cat. #5032], DNMT3A (CST, Cat. #3598), DNMT3B (CST, Cat. #72335), H3 (CST, Cat. #14269), GAPDH (CST, Cat. #2118), actin (Sigma, Cat. #A2228), and vinculin (Sigma, Cat. #SAB4200729).

Techniques: Labeling, Mass Spectrometry, Modification, Western Blot, Concentration Assay, Colony Assay

Journal: bioRxiv

Article Title: RAC1 Regulates Shh-Medulloblastoma Growth via GLI-Mediated Transcription

doi: 10.1101/2025.05.22.655563

Figure Lengend Snippet: (A-D) Representative immunoblots for RAC1, GLI1, GLI2, DNMT1 and UHRF1 levels, and their corresponding loading control (β Tubulin or β Actin) after Rac1 siRNA mediated depletion in ONS76 cells; (means ±SD, n= 3-5, p values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001), (E) Relative mRNA expression of RAC1, GLI1, GLI2, DNMT1 and UHRF1 in ONS76 cells (means ±SD, n= 2; p values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001) (Source data file).

Article Snippet: These sections were probed with primary antibody RAC1 Mouse Monoclonal antibody (Proteintech; Cat No. 66122-1-Ig), Ki-67 (Ab15580), Cleaved Caspase-3 (Asp175) Antibody (Cell signaling technology; Cat No. 9661), GLI1 Antibody (A-7) (SantaCruz Biotechnology; Cat No. sc-515781), GLI-2 Antibody (R & D Systems; Cat No. AF3635) and DNMT1 (D63A6) XP Rabbit mAb (Cell signaling technology; Cat No. 5032) Tumor tissue sections slides were subsequently incubated with VECTASTAIN® ABC-HRP Kit, Peroxidase (Mouse IgG) (PK-4002) or VECTASTAIN® ABC-HRP Kit, Peroxidase (Rabbit IgG) (PK-4001) secondary antibody.

Techniques: Western Blot, Control, Expressing

Journal: bioRxiv

Article Title: RAC1 Regulates Shh-Medulloblastoma Growth via GLI-Mediated Transcription

doi: 10.1101/2025.05.22.655563

Figure Lengend Snippet: (A) Representative immunoblots showing RAC1-GTP and RAC1 levels with their corresponding loading control (β Tubulin) after treatment with varying GYS32661 concentrations for 24 hours in ONS76 cells. (B) Immunoblot representative images showing GLI1 protein expression in ONS76 cells as compared to its corresponding loading control GAPDH and GLI1, after 8μM, GYS32661 treatment at different time points. (C-D) Immunoblot representative images showing GLI2, UHRF1, and DNMT1 expression as compared to their corresponding loading control (β Actin or GAPDH) in ONS76 cells after GYS32661 treatment at different time points. (E) Representative immunoblots showing GLI1, GLI2 and DNMT1 levels and their corresponding loading control (β Tubulin) after treatment with different concentrations of GYS32661 for 24 hours in MSC4 cells. (F) Representative immunoblots demonstrating UHRF1 levels in MSC4 cells with its corresponding loading control (β Actin) after treatment with different concentrations of GYS32661 for 24 hours. (G) Quantitative representation of relative mRNA levels of GLI1 in ONS76 cells after GYS32661 treatment at different time points (means ±SD, n=2; values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001). (H) Quantitative representation of ChIP qPCR of ONS76 cells upon GYS32661 treatment (means ±SD, n=2; values by two-way ANOVA with uncorrected Fisher’s LSD, *P<0.05, **P<0.001, ****P<0.0001 representative of 3 individual replicates); (I) Diagram representing the binding site of RAC1 at the GLI1 upstream promoter region (Source data file).

Article Snippet: These sections were probed with primary antibody RAC1 Mouse Monoclonal antibody (Proteintech; Cat No. 66122-1-Ig), Ki-67 (Ab15580), Cleaved Caspase-3 (Asp175) Antibody (Cell signaling technology; Cat No. 9661), GLI1 Antibody (A-7) (SantaCruz Biotechnology; Cat No. sc-515781), GLI-2 Antibody (R & D Systems; Cat No. AF3635) and DNMT1 (D63A6) XP Rabbit mAb (Cell signaling technology; Cat No. 5032) Tumor tissue sections slides were subsequently incubated with VECTASTAIN® ABC-HRP Kit, Peroxidase (Mouse IgG) (PK-4002) or VECTASTAIN® ABC-HRP Kit, Peroxidase (Rabbit IgG) (PK-4001) secondary antibody.

Techniques: Western Blot, Control, Expressing, ChIP-qPCR, Binding Assay

Journal: bioRxiv

Article Title: RAC1 Regulates Shh-Medulloblastoma Growth via GLI-Mediated Transcription

doi: 10.1101/2025.05.22.655563

Figure Lengend Snippet: (A & B) Representative images of PLA assays of ONS76 and MSC4 cells upon GYS32661 treatment for 24 hours showing the GLI1-DNMT1 interaction, (means ±SD, n= 5-10; values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001). (C & D) Representative images of PLA assay of ONS76 and MSC4 cells after GYS32661 treatment for 24 hours showing GLI1-UHRF1 interaction (means ±SD, n= 5-10; values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001) (Source data file).

Article Snippet: These sections were probed with primary antibody RAC1 Mouse Monoclonal antibody (Proteintech; Cat No. 66122-1-Ig), Ki-67 (Ab15580), Cleaved Caspase-3 (Asp175) Antibody (Cell signaling technology; Cat No. 9661), GLI1 Antibody (A-7) (SantaCruz Biotechnology; Cat No. sc-515781), GLI-2 Antibody (R & D Systems; Cat No. AF3635) and DNMT1 (D63A6) XP Rabbit mAb (Cell signaling technology; Cat No. 5032) Tumor tissue sections slides were subsequently incubated with VECTASTAIN® ABC-HRP Kit, Peroxidase (Mouse IgG) (PK-4002) or VECTASTAIN® ABC-HRP Kit, Peroxidase (Rabbit IgG) (PK-4001) secondary antibody.

Techniques:

Journal: bioRxiv

Article Title: RAC1 Regulates Shh-Medulloblastoma Growth via GLI-Mediated Transcription

doi: 10.1101/2025.05.22.655563

Figure Lengend Snippet: (A & B) Representative MRI images of Shh-MB tumors after in vivo MSC4 cell orthotopic injection after endpoint of the vehicle (Upper images in A) or GYS32661 (Lower images in A) treatment, and their representative H&E images, respectively; (B) Quantitation of tumor volumes of orthotopic MSC4 mouse Shh-MB tumors after the endpoint of GYS32661 cohort (means ±SD, n= 12 individual mice per group; values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001); (D) Kaplan Meier survival curve analysis of orthotopic MSC4 mouse Shh-MB tumors of GYS32661 cohort (means ±SD, n= 12 individual mice per group). (E & F) Representative IHC images of MSC4 mouse orthotopic Shh-MB tumors after the GYS32661 cohort demonstrating Ki67, Cleaved Caspase-3, GLI1, GLI2 and DNMT1 protein levels, respectively; (means ±SD, n=7 different fields of n=4 representative individual mice; values by unpaired 2-tailed t-Test *P<0.05, **P<0.001, ****P<0.0001) (Source data file).

Article Snippet: These sections were probed with primary antibody RAC1 Mouse Monoclonal antibody (Proteintech; Cat No. 66122-1-Ig), Ki-67 (Ab15580), Cleaved Caspase-3 (Asp175) Antibody (Cell signaling technology; Cat No. 9661), GLI1 Antibody (A-7) (SantaCruz Biotechnology; Cat No. sc-515781), GLI-2 Antibody (R & D Systems; Cat No. AF3635) and DNMT1 (D63A6) XP Rabbit mAb (Cell signaling technology; Cat No. 5032) Tumor tissue sections slides were subsequently incubated with VECTASTAIN® ABC-HRP Kit, Peroxidase (Mouse IgG) (PK-4002) or VECTASTAIN® ABC-HRP Kit, Peroxidase (Rabbit IgG) (PK-4001) secondary antibody.

Techniques: In Vivo, Injection, Quantitation Assay