Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting MCM6 Enhances Melphalan Chemosensitivity in Retinoblastoma by Modulating DNA Damage Response

doi: 10.1167/iovs.66.15.14

Figure Lengend Snippet: Expression of MCM6 after intravitreal melphalan chemotherapy in human retinoblastoma. ( A , B ) Immunohistochemical staining of MCM6 in human RB tissues from melphalan (Melp)-refractory cases ( A ) and percentage of MCM6 + cells compared with untreated RB tissues ( B ). Data are presented as mean ± SD ( n = 8 for untreated group; n = 6 for melphalan-refractory group), with significance assessed by unpaired two-tailed Student's t -test. ** P < 0.01.

Article Snippet: On day 14 post-injection, mice received a single intravitreal injection of 1-μL melphalan (0.25 μg/μL in PBS, HY-17575; MedChemExpress, Monmouth Junction, NJ, USA) or vehicle.

Techniques: Expressing, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting MCM6 Enhances Melphalan Chemosensitivity in Retinoblastoma by Modulating DNA Damage Response

doi: 10.1167/iovs.66.15.14

Figure Lengend Snippet: MCM6 knockdown enhances sensitivity of RB cells to melphalan. ( A , B ) Western blot analysis validating the knockdown efficiency of MCM6 in WERI-Rb1 cells. ( C ) Cell viability assays and IC 50 analysis of melphalan treatment in WERI-Rb1 and Y79 cells with or without MCM6 knockdown. Data are presented as mean ± SD ( n = 3), with IC 50 values derived from nonlinear regression (four-parameter logistic model). ( D ) Schematic diagram of the experimental design. ( E , F ) Gross and H&E staining images of enucleated eyes from different experimental groups ( E ) and quantification of tumor area in NC and shMCM6 groups without and with melphalan treatment ( F ). Data are presented as mean ± SD ( n = 5), with significance assessed by unpaired two-tailed Student's t -test. ** P < 0.01, *** P < 0.001. ( G , H ) Immunohistochemical staining of Ki67 in NC and shMCM6 groups without and with melphalan treatment ( G ) and quantification of Ki67 levels ( H ). Data are presented as mean ± SD ( n = 3), with significance assessed by unpaired two-tailed Student's t -test. **P < 0.01.

Article Snippet: On day 14 post-injection, mice received a single intravitreal injection of 1-μL melphalan (0.25 μg/μL in PBS, HY-17575; MedChemExpress, Monmouth Junction, NJ, USA) or vehicle.

Techniques: Knockdown, Western Blot, Derivative Assay, Staining, Two Tailed Test, Immunohistochemical staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Targeting MCM6 Enhances Melphalan Chemosensitivity in Retinoblastoma by Modulating DNA Damage Response

doi: 10.1167/iovs.66.15.14

Figure Lengend Snippet: MCM6 knockdown enhances melphalan sensitivity by promoting DNA damage and inhibiting the PI3K/Akt pathway. ( A , B ) Western blot analysis of phosphorylated and total mTOR, PI3K, and Akt, as well as γ-H2A.X expression, in WERI-Rb1 cells ( A ) and Y79 cells ( B ) following MCM6 knockdown and/or melphalan treatment. ( C , D ) IF staining ( C ) and quantification ( D ) of γ-H2A.X foci in WERI-Rb1 and Y79 cells under the indicated treatments. ( E , F ) Comet assay images ( E ) and quantification of tail DNA percentage ( F ) in WERI-Rb1 and Y79 cells. In D and F , data are presented as mean ± SD ( n = 3), with significance assessed by ordinary one-way ANOVA. * P < 0.05, *** P < 0.001, **** P < 0.0001; NS, not significant.

Article Snippet: On day 14 post-injection, mice received a single intravitreal injection of 1-μL melphalan (0.25 μg/μL in PBS, HY-17575; MedChemExpress, Monmouth Junction, NJ, USA) or vehicle.

Techniques: Knockdown, Western Blot, Expressing, Staining, Single Cell Gel Electrophoresis