Journal: Scientific Reports

Article Title: TMBIM6 promotes glioma progression according to integrated bioinformatics and experimental evidence

doi: 10.1038/s41598-025-06799-9

Figure Lengend Snippet: Experimental validation by immunohistochemistry and TMBIM6’s siRNAs tranfection. ( a ) TMBIM6 expression was not detected in normal brain cortex tissue, 3 of 4 lower-grade glioma (LGG) cases showed expression of TMBIM6, and 6 of 10 high-grade glioma (GBM) showed expression of TMBIM6. ( b ) Immunohistochemical analysis of TMBIM6 protein expression in Glioma compared to normal by glioma patients’ microarray slide. Healthy human Liver tissues were used as positive control. ( c ) M2 macrophage’s marker CD163 expression in Glioma compared to normal brain by immunohistochemical analysis on glioma patients’ microarray slide. Healthy human lung tissues were considered as positive control for CD163. ( d ) Quantitative real-time PCR (qPCR) analysis of TMBIM6 mRNA expression in U87-MG cells 48 h after transfection with two different TMBIM6 siRNAs and a negative control siRNA. GAPDH was used as an internal control. ( e ) Cell viability of U87-MG cells determined by MTT assay 48 h post-transfection under the same conditions as in ( d ). Data are presented as mean ± SEM of three independent experiments. Statistical significance was determined using Student’s t-test (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001).

Article Snippet: For transfection, cells were seeded in 6 cm dishes, 96 well plates and 12- well plates and grown to approximately 50–70% confluency. siRNAs targeting TMBIM6 (two distinct sequences) and siRNA negative control, along with miRNA reagents including miRNA negative control, miR-128 mimic, miR-128-3p inhibitor, and miR-128-5p inhibitor, were obtained from Bioneer Corporation, Daejeon, Republic of Korea (Order no.: Q-250403-0040).

Techniques: Biomarker Discovery, Immunohistochemistry, Expressing, Immunohistochemical staining, Microarray, Positive Control, Marker, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Control, MTT Assay

Journal: PLoS ONE

Article Title: A Role for Methyl-CpG Binding Domain Protein 2 in the Modulation of the Estrogen Response of pS2 / TFF1 Gene

doi: 10.1371/journal.pone.0009665

Figure Lengend Snippet: Real time RT-PCR analysis of pS2 transcripts in HeLa and MBD2 -depleted HeLa cells (HeLa cells pretreated for 72 h with MBD2 siRNA) transfected with an MBD2 vector expressing a transcript resistant to RNAi (pRev-MBD2 vector) or with an empty basic vector pGL3. Transcriptional expression of pS2 was analyzed 24 h after transfection. The fold change of pS2 expression was calculated from the relative pS2 mRNA in pRev-MBD2-transfected cells compared to pGL3-transfected cells. Values are presented as the mean ± standard deviation of at least three independent transfection experiments. A significant difference between the two cell groups is represented by an asterisk * (P<0.05).

Article Snippet: siRNA duplexes for MBD2 (sense: 5′-GGAGGAAGUGUACCGAAATT-3′ ; antisense: 5′-UUUUCGGAUCACUUCCUCCTT-3′ ) and non-specific siRNA control were obtained from Eurogentec (Eurogentec, Seraing, Belgium).

Techniques: Quantitative RT-PCR, Transfection, Plasmid Preparation, Expressing, Standard Deviation

Journal: PLoS ONE

Article Title: A Role for Methyl-CpG Binding Domain Protein 2 in the Modulation of the Estrogen Response of pS2 / TFF1 Gene

doi: 10.1371/journal.pone.0009665

Figure Lengend Snippet: ( A ) Transcriptional expression of pS2 was analyzed using relative RT-PCR in HeLa cells expressing ERα and/or depleted in MBD2. Mock, mock-treated HeLa cells. ER, HeLa cells 24 h after transfection with a human ERα expression vector, HEG0. MBD2 siRNA, HeLa cells pretreated for 72 h with MBD2 siRNA and again for 24 h. MBD2 siRNA + ER, HeLa cells pretreated for 72 h with MBD2 siRNA, then cotransfected with MBD2 siRNA and HEG0 for 24 h. OHT, 24 h treatment with 4-hydroxytamoxifen. MCF7, MCF7 cells. ( B ) Bar chart showing the fold change of pS2 expression in HeLa cells expressing ERα and/or depleted in MBD2. pS2 transcripts were quantified by real time RT-PCR. The fold change was calculated from the relative pS2 mRNA in treated compared to mock-treated cells. Each bar represents the mean ± standard deviation of three analyses. A significant difference between the two cell groups is represented by an asterisk * (P<0.05).

Article Snippet: siRNA duplexes for MBD2 (sense: 5′-GGAGGAAGUGUACCGAAATT-3′ ; antisense: 5′-UUUUCGGAUCACUUCCUCCTT-3′ ) and non-specific siRNA control were obtained from Eurogentec (Eurogentec, Seraing, Belgium).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Quantitative RT-PCR, Standard Deviation