Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: PROTAC Mcl-1 Degrader-1 reduced the viability of HT-29 and COLO-205 cells. ( A ) Molecular structure of PROTAC Mcl-1 Degrader-1 (The pink end of the PROTAC molecule binds to the target protein, while the blue end binds to the E3 ligase). Cell viability of ( B , C ) Colorectal cancer cell lines (HT-29/COLO-205) was determined by WST-8 assay after 24 and 48 h of treatment with PROTAC Mcl-1 Degrader-1. (n = 3 replicates). ( D , E ) In a 48 h combination therapy where trametinib remained stable at 1 µM and PROTAC Mcl-1 Degrader-1 doses varied, it was shown to effectively reduce the viability of ( D ) HT-29 and ( E ) COLO-205 cells. ( F ) Western blot analysis showed a decrease in MCL-1 protein levels in PROTAC Mcl-1 Degrader-1 colorectal cancer cell lines. Three repetitions were performed in each group. ( G ) Mcl-1 and Bcl-2 protein expressions were statistically evaluated. The results were obtained by one-way ANOVA test * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and ‘ns’ means not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Western Blot

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: PROTAC Mcl-1 Degrader-1 and Trametinib affect the proliferation of HT-29 and COLO-205 cells. ( A – C ) Simultaneous results obtained by Xcelligence Real-Time Proliferation assay after 48 h of treatment of HT-29 cells with ( A ) PROTAC Mcl-1 Degrader-1 and ( B ) Trametinib. Doses are represented by the same colors in the graph. ( D – F ) Proliferation results of COLO-205 cell line after 48 h of single and combined treatment with ( D ) PROTAC Mcl-1 Degrader-1 and ( E ) Trametinib, respectively. In combination, a dose of 1 µM Trametinib was used, and treatment was applied with variable doses of PROTAC Mcl-1 Degrader-1.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Proliferation Assay

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: Effect of PROTAC Mcl-1 Degrader-1 on the migratory ability of colorectal cancer cells. ( A ) Wound healing assay was performed under a microscope (original magnification 4×) using an inverted microscope (500 µm scale bar) and images were taken at 0, 24 and 48 time intervals after treatment with different doses of PROTAC Mcl-1 Degrader-1 in HT-29 and ( B ) COLO-205 cell lines. ( C , D ) Wound closure analyses of HT-29 and COLO-205 cells were measured using PROTAC Mcl-1 Degrader-1 and microscopic images taken at 24 (black bars) and 48 (red bars) time intervals. Results are presented as mean ± standard deviation (n = 3/group). Statistical significance was determined using one-way ANOVA and Tukey’s multiple comparison test (GraphPad Prism 10.0, GraphPad Software Inc.). * p < 0.05; ** p < 0.01; ns = not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Wound Healing Assay, Microscopy, Inverted Microscopy, Standard Deviation, Comparison, Software

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: The combination of PROTAC Mcl-1 Degrader-1 and Trametinib did not lead to a statistically significant change in cell migration. ( A , B ) Colorectal cancer cells HT-29 and COLO-205 were treated with a combination of PROTAC Mcl-1 Degrader-1 and Trametinib. Trametinib dose was kept constant at 1 µM, while PROTAC Mcl-1 Degrader-1 doses were varied. Cell migration analysis was performed after treatment. Migration capabilities at 0, 24, and 48 h were visualized under a microscope. ( C , D ) Graphical analysis of wound closure was performed to determine the migratory ability of cells. The analysis was performed by comparing the 24 and 48 h treatments with the control group (ns = not significant).

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Migration, Microscopy, Control

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: Effect of PROTAC Mcl-1 Degrader-1 on cell count in colorectal cancer. ( A ) Flow cytometric analysis of the preservation of HT-29 and COLO-205 cell counts after 48 h of maintenance with PROTAC Mcl-1 Degrader-1 (n = 3). ( B , C ) Statistical analysis of the percentage cell counts of HT-29 and COLO-205 cells. Statistical significance was determined using one-way ANOVA and Tukey’s multiple comparison test (GraphPad Prism 10.0, GraphPad Software Inc.). * p < 0.05; ns = not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Cell Characterization, Preserving, Comparison, Software

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: Combination treatment with PROTAC Mcl-1 Degrader-1 and Trametinib for 48 h showed a more pronounced effect on cell cycle distribution in HT-29 cells compared to COLO-205 cells. ( A ) A graph of the cell cycle percentage of HT-29 and COLO-205 cells was plotted after the combination. ( B , C ) As shown in the statistical analysis, an effect was observed in the S and G2/M phases in the HT-29 cell line, while no significant effect was statistically demonstrated in the COLO-205 cell line. Statistical significance was determined using one-way ANOVA and Tukey’s multiple comparison test (GraphPad Prism 10.0, GraphPad Software Inc.). * p < 0.05; ns = not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Comparison, Software

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: The combination of PROTAC Mcl-1 Degrader-1 and Trametinib affected mitochondrial membrane potential. ( A ) HT-29 and COLO-205 cell lines were stained with JC-1 after treatment with PROTAC Mcl-1 Degrader-1 for 48 h and imaged under a fluorescence microscope (original magnification 20×). The yellow-orange fluorescence of JC-1 dimers was found in cell regions with high mitochondrial membrane potential, while the green fluorescence of JC-monomers was prevalent in cell regions with low mitochondrial membrane potential. ( B , C ) Percentage red/green fluorescence intensity plots showed that HT-29 and COLO-205 cells were converted to MMP in both cell lines after 48 h of treatment with PROTAC Mcl-1 Degrader-1, particularly at the final dose of 10 μM. ( D ) Cell lines treated with a combination of PROTAC Mcl-1 Degrader-1 and Trametinib at different 48 h doses were subjected to JC-1 staining. After staining (original magnification 20×), images were acquired under a fluorescence microscope. ( E , F ) Quantitative analysis of the transition from mitochondrial orange to green coloration between different combination treatment groups. The statistical value of this experiment was determined using one-way ANOVA and Tukey’s multiple comparison test (GraphPad Prism 10.0, GraphPad Software Inc.). ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns = not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Membrane, Staining, Fluorescence, Microscopy, Comparison, Software

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: PROTAC Mcl-1 Degrader-1 alone was associated with a partial increase in apoptotic cell populations. (Q1 LL (Control), Q1 UL (Necrosis), Q1 UR (Late apoptosis), Q1 LR (Early apoptosis). ( A ) After 48 h of PROTAC Mcl-1 Degrader-1 treatment of HT-29 and COLO-205 cells, Annexin V and PI-labeled cell apoptosis was analyzed by flow cytometry. ( B , C ) Statistical analysis of cells in ( B ) early and ( C ) late apoptosis phases of HT-29 cell line after treatment with PROTAC Mcl-1 Degrader-1. ( D , E ) Statistical analysis of COLO-205 cells in ( D ) early and ( E ) late apoptosis phases after treatment with PROTAC Mcl-1 Degrader-1. The obtained results were presented using mean ± standard deviation (n = 3/group). Statistical significance in the experiment was determined using one-way ANOVA and Tukey’s test for multiple comparisons (GraphPad Prism 10.0, GraphPad Software Inc.). * p < 0.05.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Control, Labeling, Flow Cytometry, Standard Deviation, Software

Journal: Current Issues in Molecular Biology

Article Title: Targeted Degradation of MCL-1 by PROTAC Mcl-1 Degrader-1 Exhibits Antiproliferative and Antimigratory Effects and Triggers Mitochondria-Mediated Apoptosis in Colorectal Cancer

doi: 10.3390/cimb48070733

Figure Lengend Snippet: The combination of PROTAC Mcl-1 Degrader-1 and trametinib was observed to be associated with an increase in apoptotic cell populations. (Q1 LL (Control), Q1 UL (Necrosis), Q1 UR (Late apoptosis), Q1 LR (Early apoptosis). ( A ) The degree of apoptosis in HT-29 and COLO-205 cells was monitored by flow cytometry after 48 h of exposure to a combination of PROTAC Mcl-1 Degrader-1 and Trametinib. Combination therapy was shown to be associated with a higher level of apoptosis compared to monotherapy. ( B , C ) Treatment of the HT-29 cell line with the combination of PROTAC Mcl-1 Degrader-1 and Trametinib showed increased early and late apoptosis graphs, indicating the stages of apoptosis, compared to the control group. ( D , E ) The outcome of combination therapy in the COLO-205 cell line shows a significant effect on early apoptosis in the early and late apoptosis graphs. Statistical analysis after combination therapy showed that the final dose of 1 µm Trametinib and 10 µm PROTAC Mcl-1 Degrader-1 increased early apoptosis by 23% and late apotosis by 8%, particularly in the COLO-205 cell line. The values are expressed as the mean ± SD (n = 3/group). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparison test. * p < 0.05; ** p < 0.01; ns = not significant.

Article Snippet: PROTAC Mcl-1 Degrader-1 (compound C3 ) (Catalog number: HY-125877) and Trametinib (Catalog number: HY-10999) were purchased from MedChem Express (Township, NJ, USA) and dissolved in DMSO.

Techniques: Control, Flow Cytometry, Comparison

Journal: bioRxiv

Article Title: Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy

doi: 10.64898/2026.05.06.722533

Figure Lengend Snippet: Bifunctional small molecule dTAG-13 binds to the FKBP12 F36V domain of tagged CYP1A2 and recruits an E3 ligase; ubiquitination (curved arrow) directs fusion protein degradation. The AP1867 moiety of dTAG-13 (bottom left) binds FKBP12 F36V selectively; the thalidomide moiety recruits cereblon.

Article Snippet: dTAG-13 was purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Ubiquitin Proteomics

Journal: bioRxiv

Article Title: Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy

doi: 10.64898/2026.05.06.722533

Figure Lengend Snippet: a , Hepatic CYP1A2 and CYP1A2 dTAG protein expression in WT, HET, and HOM mice. b , Densitometric quantification of CYP1A2 and CYP1A2 dTAG protein. c , CYP1A2 dTAG catalytic activity. d , Dose-dependent degradation of CYP1A2 dTAG protein mediated by dTAG-13 in HMO mice. e , Densitometric quantification of CYP1A2 dTAG protein in liver 4 hours post dTAG-13 treatment. f & g , CYP1A2 dTAG activity in MLMs prepared from HOM mice treated with vehicle or dTAG-13. h , Hepatic immunostaining of CYP1A2 dTAG in HOM mice (vehicle). i , Hepatic immunostaining of CYP1A2 dTAG in HOM mice (dTAG-13). For densitometric quantification in , native Cyp1a2 level was set as 100%; in , protein levels in vehicle-treated mice were set as 100%. M, male; F, Female. na, not available; ns, not significant; **, p<0.01, ***; P<0.001; HET, heterozygous; HOM, homozygous. n = 4.

Article Snippet: dTAG-13 was purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: Expressing, Activity Assay, Immunostaining

Journal: bioRxiv

Article Title: Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy

doi: 10.64898/2026.05.06.722533

Figure Lengend Snippet: a , HOM-v vs. WT-v. b , HOM-d vs. HOM-v. v, vehicle; d, dTAG-13. * Cyp1a2 in the figure refers to CYP1A2 or CYP1A2 dTAG . (n = 6, 3 males and 3 females).

Article Snippet: dTAG-13 was purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques:

Journal: bioRxiv

Article Title: Reversible in vivo regulation of drug metabolizing enzyme CYP1A2 activity through a dTAG knock-in strategy

doi: 10.64898/2026.05.06.722533

Figure Lengend Snippet: a , The PK profile of caffeine. b , The PK profile of paraxanthine (a CYP1A2-mediated primary metabolite). c , The PK parameters of caffeine. The caffeine (10 mg/kg, p.o.) was administered to mice 4 h post vehicle or dTAG-13 (20 mg/kg, i.p.). Two-week wash-out following PK of caffeine in Cyp1a2 dTAG -HOM mice treated with vehicle, PK in Cyp1a2 dTAG -HOM mice treated with dTAG-13 was performed. (n = 6). *, p<0.05; **, p<0.01, ***. P<0.001. HOM, homozygous.

Article Snippet: dTAG-13 was purchased from MedChemExpress (Monmouth Junction, NJ).

Techniques: