1 ap  (Proteintech)

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: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies, including p-STAT3 (cat. no. 4113, Cell Signaling Technology), MCL1 (cat. no. 16225-1-AP), MMP2 (cat. no. 10373-2-AP) and SOX2 (cat. no. 11064-1-AP; all Proteintech Group, Inc.), were diluted 1:100 in antibody diluent (cat. no. PR30016; Proteintech Group, Inc.) and applied at 4°C overnight.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing