Journal: Reports of Biochemistry & Molecular Biology

Article Title: The Prognostic Significance of P16 Immunohistochemical Expression Pattern in Women with Invasive Ductal Breast Carcinoma

doi: 10.52547/rbmb.12.1.83

Figure Lengend Snippet: Immunohistochemical staining for p16 protein in invasive ductal breast tumors. A. p16 negative, B. p16 low positive, C. High-positive.

Article Snippet: Parameters P16 Protein Expression Level P-value(Chi-Square) Negative (n = 29) Low-Positive (n = 44) High-Positive (n = 27) ER receptor Positive 22 (75.9%) 41 (93.2%) 17 (63%) 0.007* Negative 7 (24.1%) 3 (6.8%) 10 (37%) PR receptor Positive 14 (48.3%) 33 (75%) 16 (59.3%) 0.061 Negative 15 (51.7%) 11 (25%) 11 (40.7%) HER2 Positive 4 (13.8%) 15 (34.1%) 11 (40.7%) 0.065 Negative 25 (86.2%) 29 (65.9%) 16 (59.3%) Ki67 Positive (> 14%) 7 (24.1%) 16 (36.4%) 5 (18.5%) 0.229 Negative (≤ 14%) 22 (75.9%) 28 (63.6%) 22 (81.5%) Cancer Grade I 2 (6.9%) 2 (4.5%) 4 (14.8%) 0.252 II 20 (69%) 37 (84.1%) 17 (6%) III 7 (24.1%) 5 (11.4%) 6 (22.2%) Tumor Size ≤ 20 mm 11 (37.9%) 18 (40.9%) 13 (48.1%) 0.754 21-50 mm 18 (62.1%) 25 (56.8%) 14 (51.9%) > 50 mm 0 (0%) 1 (2.3%) 0 (0%) Cancer Type Luminal A 7 (29.16%) 13 (54.16%) 4 (16.66%) 0.144 Luminal B 15 (26.31%) 28 (49.12%) 14 (24.56%) HER2 positive 2 (28.57%) 1 (14.28%) 4 (57.14%) Triple Negative 5 (41.66%) 2 (16.66%) 5 (41.66%) Age ≤ 40 years 7 (24.1%) 11 (25%) 10 (37%) 0.471 > 40 yea 22 (75.9%) 33 (75%) 17 (63%) Family History Yes 5 (17.2%) 11 (25%) 5 (18.5%) 0.68 No 24 (82.8%) 33 (75%) 22 (81.5%) Open in a separate window The data are shown as mean ± SD.

Techniques: Immunohistochemical staining, Staining

Journal: Reports of Biochemistry & Molecular Biology

Article Title: The Prognostic Significance of P16 Immunohistochemical Expression Pattern in Women with Invasive Ductal Breast Carcinoma

doi: 10.52547/rbmb.12.1.83

Figure Lengend Snippet: Correlation between clinicopathological characteristics and p16 protein expression in women with invasive ductal breast carcinoma.

Article Snippet: Parameters P16 Protein Expression Level P-value(Chi-Square) Negative (n = 29) Low-Positive (n = 44) High-Positive (n = 27) ER receptor Positive 22 (75.9%) 41 (93.2%) 17 (63%) 0.007* Negative 7 (24.1%) 3 (6.8%) 10 (37%) PR receptor Positive 14 (48.3%) 33 (75%) 16 (59.3%) 0.061 Negative 15 (51.7%) 11 (25%) 11 (40.7%) HER2 Positive 4 (13.8%) 15 (34.1%) 11 (40.7%) 0.065 Negative 25 (86.2%) 29 (65.9%) 16 (59.3%) Ki67 Positive (> 14%) 7 (24.1%) 16 (36.4%) 5 (18.5%) 0.229 Negative (≤ 14%) 22 (75.9%) 28 (63.6%) 22 (81.5%) Cancer Grade I 2 (6.9%) 2 (4.5%) 4 (14.8%) 0.252 II 20 (69%) 37 (84.1%) 17 (6%) III 7 (24.1%) 5 (11.4%) 6 (22.2%) Tumor Size ≤ 20 mm 11 (37.9%) 18 (40.9%) 13 (48.1%) 0.754 21-50 mm 18 (62.1%) 25 (56.8%) 14 (51.9%) > 50 mm 0 (0%) 1 (2.3%) 0 (0%) Cancer Type Luminal A 7 (29.16%) 13 (54.16%) 4 (16.66%) 0.144 Luminal B 15 (26.31%) 28 (49.12%) 14 (24.56%) HER2 positive 2 (28.57%) 1 (14.28%) 4 (57.14%) Triple Negative 5 (41.66%) 2 (16.66%) 5 (41.66%) Age ≤ 40 years 7 (24.1%) 11 (25%) 10 (37%) 0.471 > 40 yea 22 (75.9%) 33 (75%) 17 (63%) Family History Yes 5 (17.2%) 11 (25%) 5 (18.5%) 0.68 No 24 (82.8%) 33 (75%) 22 (81.5%) Open in a separate window The data are shown as mean ± SD.

Techniques: Expressing

Journal: Reports of Biochemistry & Molecular Biology

Article Title: The Prognostic Significance of P16 Immunohistochemical Expression Pattern in Women with Invasive Ductal Breast Carcinoma

doi: 10.52547/rbmb.12.1.83

Figure Lengend Snippet: Correlation of p16 expression with tumor grade and age of patients with invasive ductal breast carcinoma. The data are shown as mean ± SD.

Article Snippet: Parameters P16 Protein Expression Level P-value(Chi-Square) Negative (n = 29) Low-Positive (n = 44) High-Positive (n = 27) ER receptor Positive 22 (75.9%) 41 (93.2%) 17 (63%) 0.007* Negative 7 (24.1%) 3 (6.8%) 10 (37%) PR receptor Positive 14 (48.3%) 33 (75%) 16 (59.3%) 0.061 Negative 15 (51.7%) 11 (25%) 11 (40.7%) HER2 Positive 4 (13.8%) 15 (34.1%) 11 (40.7%) 0.065 Negative 25 (86.2%) 29 (65.9%) 16 (59.3%) Ki67 Positive (> 14%) 7 (24.1%) 16 (36.4%) 5 (18.5%) 0.229 Negative (≤ 14%) 22 (75.9%) 28 (63.6%) 22 (81.5%) Cancer Grade I 2 (6.9%) 2 (4.5%) 4 (14.8%) 0.252 II 20 (69%) 37 (84.1%) 17 (6%) III 7 (24.1%) 5 (11.4%) 6 (22.2%) Tumor Size ≤ 20 mm 11 (37.9%) 18 (40.9%) 13 (48.1%) 0.754 21-50 mm 18 (62.1%) 25 (56.8%) 14 (51.9%) > 50 mm 0 (0%) 1 (2.3%) 0 (0%) Cancer Type Luminal A 7 (29.16%) 13 (54.16%) 4 (16.66%) 0.144 Luminal B 15 (26.31%) 28 (49.12%) 14 (24.56%) HER2 positive 2 (28.57%) 1 (14.28%) 4 (57.14%) Triple Negative 5 (41.66%) 2 (16.66%) 5 (41.66%) Age ≤ 40 years 7 (24.1%) 11 (25%) 10 (37%) 0.471 > 40 yea 22 (75.9%) 33 (75%) 17 (63%) Family History Yes 5 (17.2%) 11 (25%) 5 (18.5%) 0.68 No 24 (82.8%) 33 (75%) 22 (81.5%) Open in a separate window The data are shown as mean ± SD.

Techniques: Expressing

Journal: Breast Cancer Research : BCR

Article Title: Evaluating the predictive value of biomarkers for efficacy outcomes in response to pertuzumab- and trastuzumab-based therapy: an exploratory analysis of the TRYPHAENA study

doi: 10.1186/bcr3690

Figure Lengend Snippet: The relationship between biomarker levels and whether a pathologic complete response (pCR) was achieved, adjusted for estrogen receptor status (all arms pooled)

Article Snippet: Targos Molecular Pathology GmbH, Kassel, Germany conducted analysis of: mRNA expression levels of HER2 , HER3, EGFR, amphiregulin, and betacellulin ; protein expression levels of HER3, EGFR, IGF-1R, and PTEN and of c-myc and TOP2A gene amplification.

Techniques: Biomarker Discovery, Membrane, Mutagenesis

Journal: Breast Cancer Research : BCR

Article Title: Evaluating the predictive value of biomarkers for efficacy outcomes in response to pertuzumab- and trastuzumab-based therapy: an exploratory analysis of the TRYPHAENA study

doi: 10.1186/bcr3690

Figure Lengend Snippet: Comparison of baseline levels of biomarkers derived from tissue samples with the levels detected at surgery. (A) PTEN nuc, (B) HER2-CR, ( C ) EGFR-CR, (D) HER2-mem. (In the box plot, the horizontal line represents the median value, the diamond represents the mean, the upper and lower bounds of the box represent the 75th and 25th quartiles, respectively, and whiskers represent 95% confidence limits). CR, concentration ratio; EGFR, epidermal growth factor receptor; HER2, human epidermal growth factor receptor 2; mem, membrane; nuc, nuclear; PTEN, phosphatase and tensin homolog.

Article Snippet: Targos Molecular Pathology GmbH, Kassel, Germany conducted analysis of: mRNA expression levels of HER2 , HER3, EGFR, amphiregulin, and betacellulin ; protein expression levels of HER3, EGFR, IGF-1R, and PTEN and of c-myc and TOP2A gene amplification.

Techniques: Comparison, Derivative Assay, Concentration Assay, Membrane

Journal: Biomolecules

Article Title: Differential MicroRNA Expression Involved in Endometrial Receptivity of Goats

doi: 10.3390/biom11030472

Figure Lengend Snippet: Chi-miR-483 targets the 3′-untranslated region (UTR) of deltex E3 ubiquitin ligase 3L (DTX3L). ( A ) The predicted binding site of chi-miR-483 in the 3′UTR of DTX3L according to bioinformatics analysis. ( B ) Design of the luciferase reporter. WT, the wildtype sequence of DTX3L-3′UTR contains the chi-miR-483 binding site; Mut, the sequence of DTX3L-3′UTR with a mutation in the chi-miR-483 binding site. ( C ) 293T cells were co-transfected with wildtype (WT) or mutant (Mut) luciferase reports of DTX3L 3′UTR with chi-miR-483 mimics or negative control (NC) mimics. The luciferase reporter assay demonstrated that chi-miR-483 significantly decreased the luciferase activity of DTX3L WT in 293T cells. Data are shown as the mean ± SEM values ( n = 3, ** p < 0.01, Student’s t -test).

Article Snippet: Differences in wildtype (WT) or mutant (Mut) DTX3L 3′UTR luciferase reports and the expression level of DTX3L protein under two conditions were compared using Student’s t -test (GraphPad Prism version 8.0, San Diego, CA, USA).

Techniques: Ubiquitin Proteomics, Binding Assay, Luciferase, Sequencing, Mutagenesis, Transfection, Negative Control, Reporter Assay, Activity Assay

Journal: Biomolecules

Article Title: Differential MicroRNA Expression Involved in Endometrial Receptivity of Goats

doi: 10.3390/biom11030472

Figure Lengend Snippet: Immunohistochemical analysis of DTX3L in the C16 and P16 uterus. ( A ) Images stained with DTX3L antibodies. The positive signal of DTX3L was distinctly detected in the uterine luminal epithelium or glandular epithelium in P16. The section stained with nonrelevant immunoglobulin G served as the negative control (NC). ( B ) Quantitative analysis of DTX3L by measuring the average integrated optical density (IOD) in the endometrium. Asterisks indicate significant differences (mean ± SEM) between C16 and P16 (*** p < 0.001); the p -value was determined by Student’s t -test. Legend: LE, endometrial luminal epithelium; GE, glandular epithelium. Scale bar = 100 μm.

Article Snippet: Differences in wildtype (WT) or mutant (Mut) DTX3L 3′UTR luciferase reports and the expression level of DTX3L protein under two conditions were compared using Student’s t -test (GraphPad Prism version 8.0, San Diego, CA, USA).

Techniques: Immunohistochemical staining, Staining, Negative Control

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A In silico TMBIM6 mRNA expression in human CNS from THPA database. B Tmbim6 mRNA expression on N2a cells after 18 h of exposure to 25 μM 6-OHDA or 50 μM rotenone. C Tmbim6 mRNA levels over time in PCNs exposed to aSyn for 96 h. D Changes of expression of Tmbim6 , BcL2 , and Bax over time in PCNs exposed to aSyn for 96 h. E Representative Western blots of total protein extracts from postmortem human SN from neurologically healthy controls and PD patients, probed for TMBIM6 and GAPDH (loading control). Full, uncropped blots are provided in Supplementary Material. F Densitometric quantification of TMBIM6 from blots in ( E ). Band intensities were normalized to GAPDH for each lane; individual data points are shown with mean ± SEM (n = 9–10 per group). For experiments involving more than two groups, a two-way ANOVA followed by Tukey’s multiple comparison test was performed. Pairwise comparisons between two groups were analyzed using unpaired t-test ( B ) or the Mann–Whitney U test ( F ). All bars represent mean ± SEM. Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; **=p < 0.01; ***=p < 0.001; ****=p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: In Silico, Expressing, Western Blot, Control, Comparison, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A Graph shows validation of decreased Tmbim6 mRNA levels after 48 h of siRNA transfection in SN4741. B , C Representative immunoblots and quantification show mTmbim6 KD cells after 48 h. D Cytotoxicity assay shows cell death induced by 10 μM Tunicamycin after 24 h in KD cells. Results are expressed as % of LDH release. E Cytotoxicity assay shows cell death induced by 50 μM 6-OHDA after 24 h in KD cells. F Retention of DiOC6(3) assay shows the effect of aSyn on ΔΨm in KD cells after 18 h. Results are expressed as % of DiOC6(3) retention. G MTT assay shows mitochondrial-dependent cell death induced by aSyn in KD cells after 24 h. Results are expressed as % of MTT. H DEVD-AMC fluorescent assay shows the effect of aSyn on Caspase-3 activity in Tmbim6 KD cells after 24 h. Results are expressed as fold change of DEVD-AMC fluorescence intensity relative to vehicle. I Cytotoxicity assay shows the KD cell death induced by aSyn after 24 h. J Representative immunoblot of high–molecular-weight (HMW) aSyn species in SN4741 cells transfected with si Cntrl or si Tmbim6 and treated with 10 µM aSyn PFFs for 24 h; Tubulin was used as a loading control. K Densitometric quantification of HMW aSyn bands from the experiment described in ( J ) (integrated density normalized to Tubulin). All bars represent mean ± SEM. For experiments involving more than two groups, a two-way ANOVA followed by Tukey’s multiple comparison test was performed. Pairwise comparisons between two groups were analyzed using unpaired t-test ( A , C ) or the Mann–Whitney U test ( K ). Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; **=p < 0.01; ***=p < 0.001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: Biomarker Discovery, Transfection, Western Blot, Cytotoxicity Assay, MTT Assay, Fluorescence, Activity Assay, High Molecular Weight, Control, Comparison, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A d Tmbim6 mRNA expression on homogenized flies’ heads. Results are expressed as fold change of mRNA expression. The bars represent mean ± SEM. B Optic image of eye integrity in RNAi-dTmbim6 flies incubated at 25 °C. C Quantification of eye integrity score in RNAi- dTmbim6 flies incubated at 25 °C (n = 25 per group). D Immunostaining of TH+ neurons in the lamina of RNAi-dTmbim6 flies incubated at 25 °C (n = 6 per group). E Quantification of the number of TH+ neurons in the lamina of RNAi-dTmbim6 flies incubated at 25 °C (n = 6 per group). F Schematic representation of rotenone-induced PD model in D. mel . G Spontaneous activity in DAergic RNAi-dTmbim6 flies after exposition to rotenone 300 μM for 7 days (n = 6 populations of 12 flies). H Climbing assay showed the motor ability of DAergic RNAi-dTmbim6 flies exposed to rotenone 300 μM for 7 days (n = 12 per group). I Representative confocal images of IF assay showed TH+ cells from DAergic RNAi-dTmbim6 flies exposed to rotenone 300 μM for 7 days. J The numbers of TH+ cells were quantified in each DAergic cluster, and K the somal size was analyzed (n = 7 per group). All bars represent mean ± SEM. For experiments involving more than two groups, a two-way ANOVA followed by Tukey’s multiple comparison test was performed. Pairwise comparisons between two groups were analyzed using unpaired t-test ( A , C ) or the Mann–Whitney U test ( E ). Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; ** = p < 0.01; ***= p < 0.001; ****= p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: Expressing, Incubation, Immunostaining, Activity Assay, Climbing Assay, Comparison, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A Representative immunoblot shows a stable expression of TMBIM6 HA in SN4741 cells. B Cytotoxicity assay shows the cell death induced by Tunicamycin in SN4741 TMBIM6 HA cells after 24 h. C , D MTT assay shows cell death induced by 6-OHDA or rotenone in SN4741 TMBIM6 HA cells after 24 h. Results are expressed as % of MTT. E Retention of DiOC6(3) assay shows the effect of aSyn on ΔΨm in SN4741 hTMBIM6 HA cells after 18 h. Results are expressed as % of DiOC6(3) retention. F MTT assay shows cell death induced by aSyn in SN4741 TMBIM6 HA cells after 24 h. G DEVD-AMC fluorescent assay shows the effect of aSyn on Caspase-3 activity in SN4741 TMBIM6 HA cells after 24 h. H Cytotoxicity assay shows the cell death induced by aSyn in SN4741 TMBIM6 HA cells after 24 h. I Representative immunoblot of HMW aSyn species in Mock or TMBIM6 HA cells treated with aSyn PFFs for 24 h; TCE staining was used as a loading control. J Densitometric quantification of HMW aSyn bands from the experiment described in J (integrated density normalized to TCE). K Representative immunoblot shows expression of TMBIM6 HA and TMBIM6 D213A/HA in SN4741 cells. L MTT assay shows cell death induced by Tunicamycin and Thapsigargin in SN4741 Mock, TMBIM6 HA , and TMBIM6 D213A/HA cells after 24 h. Results are expressed as % of MTT. M Cytotoxicity assay shows the cell death induced by aSyn in SN4741 TMBIM6 HA and TMBIM6 D213A/HA cells after 24 h. N Cytotoxicity assay shows the cell death induced by aSyn after 10 days in PCNs transfected with Mock, TMBIM6 HA and TMBIM6 D213A/HA constructs. All bars represent mean ± SEM. For experiments involving more than two groups, a two-way ANOVA followed by Tukey’s multiple comparison test was performed. Pairwise comparisons between two groups were analyzed using the Mann–Whitney U test. Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; **=p < 0.01; ***=p < 0.001; ****=p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: Western Blot, Expressing, Cytotoxicity Assay, MTT Assay, Fluorescence, Activity Assay, Staining, Control, Transfection, Construct, Comparison, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A An in-silico assay using Ingenuity Pathway Analysis (IPA) software shows the canonical pathways significantly associated with TMBIM6 interactors. B , C UMAP visualizations of the snRNA-seq dataset (GEO: GSE178265 ) from human postmortem substantia nigra. B shows TMBIM6 expression across all nuclei, while C distinguishes nuclei from healthy and PD donors. D Dot plot comparing the expression of TMBIM6 and UPR-related genes ( HSPA5, ERN1, XBP1, BLOC1S1 ) between healthy and PD conditions across all nuclei. E UMAP plot identifying resistant and vulnerable DAergic neuron populations within the dataset. F Dot plot comparing gene expression between resistant and vulnerable DAergic neurons within the PD cohort. For dot plots ( D , F ), dot size represents the percentage of cells expressing the gene, and color intensity indicates the mean expression level. Statistical significance for the differential expression shown in ( D , F ) was determined using the Model-based Analysis of Single-cell Transcriptomics (MAST) test. Full statistical details, including FDR-adjusted p-values, are provided in Supplementary Fig. .

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: In Silico, Software, Expressing, Gene Expression, Quantitative Proteomics, Single-cell Transcriptomics

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A Representative images show red fluorescent dots of PLA assay to TMBIM6 HA /IRE1a in stable SN4741 TMBIM6 HA cells exposed to aSyn. B Quantification of PLA dots per cell. Kruskal-Wallis followed by Dunn’s multiple comparison test. C In SN4741 siRNA- mTMBIM6 cells, an RT-qPCR assay shows the effect of aSyn on mRNA levels of mouse XBP1s ( mXbp1s ), D mouse BLOCS1 (mBlocs1) , and E mouse BIP (mBip) . F In SN4741 TMBIM6 HA cells, RT-qPCR assay shows the effect of aSyn in mXbp1s , G mBlocs1 , and H mBip , mRNA levels. Results are expressed as fold change, and bars represent mean ± SEM. All bars represent mean ± SEM. In ( B ), a Kruskal–Wallis test was performed followed by Dunn’s multiple comparison test. For experiments involving more than two groups, a two-way ANOVA followed by Tukey’s multiple comparison test was performed ( C – H ). Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; ***=p < 0.001; ****=p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: Comparison, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A , B Cytotoxicity assay shows the effect of IRE1a inhibition using MKC or 4 μ 8c on cell death induced by aSyn in Tmbim6 KD cells after 24 h. Results are expressed as % of LDH release. C Cytotoxicity assay shows the effect of PERK inhibitor on cell death induced by aSyn in mTmbim6 KD cells after 24 h. D A RT-qPCR assay shows effective double knockdown of both mTmbim6 (left panel) and mIre1 a (right panel) in SN4741 cells. Results are expressed as fold change, and bars represent mean ± SEM. One way ANOVA, Dunnett´s multiple comparation test. E Cytotoxicity assay shows downregulation of mIRE1a over cell death induced by aSyn in mTmbim6 KD cells after 24 h. Results are expressed as % of LDH release. F Cytotoxicity assay shows the JNK inhibitor AS60125 over cell death induced by aSyn in mTmbim6 KD cells after 24 h. G Cytotoxicity assay shows the BAX inhibitor BAI-1 over cell death induced by aSyn in mTmbim6 KD cells after 24 h. H Cytotoxicity assay shows the pan-caspase inhibitor ZVAD-FMK (casp-inh) over cell death induced by aSyn in mTmbim6 KD cells after 24 h. All bars represent mean ± SEM. In ( D ), a one-way ANOVA followed by Dunnett’s multiple comparison test was performed, whereas in ( E , F , G , H ), a two-way ANOVA with Tukey’s multiple comparisons test was conducted. Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; **=p < 0.01; ***=p < 0.001; ****=p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: Cytotoxicity Assay, Inhibition, Quantitative RT-PCR, Knockdown, Comparison

Journal: Cell Death & Disease

Article Title: TMBIM6 enhances dopaminergic neuron survival by modulating the IRE1a pathway in Parkinson’s disease

doi: 10.1038/s41419-025-08391-5

Figure Lengend Snippet: A Effect of two 6-OHDA doses on motor performance of mice, as a pharmacologic in vivo PD model (n = 4 per condition). Results are expressed as a percentage of contralateral forelimb use. B Timeline of in vivo transduction of the AAV-TMBIM6 HA/GFP and AAV-Mock GFP in SN and motor performance measurements in mice wild-type lesioned with 6-OHDA in CPu. C Cylinder test shows the effect of AAV-TMBIM6 HA/GFP and AAV-Mock GFP expression in SN over forelimb use after 2-, 3-, 4-, and 5-weeks post-injection (wpi). The colored area shows treatment with 6-OHDA injuries in the CPu. Results are expressed as a percentage of contralateral forelimb use (n Mockl = 4 and n TMBIM6 = 5). D Beam test shows the effect of the AAV-TMBIM6 HA/GFP and AAV-Mock GFP expression in SN on balance and coordination after 2, 3, 4, and 5 weeks after injection. The colored area shows treatment with 6-OHDA injury in the CPu. Results are expressed as the number of paws slips (n Mockl = 4 and n TMBIM6 = 5). E Effect of the AAV-TMBIM6 HA/GFP and AAV-Mock GFP expression over time, animals used to complete the Beam test during 2-, 3-, 4-, and 5-wpi. The colored area shows treatment with 6-OHDA injury in the CPu. Results are expressed as the time in seconds to complete the test (n Mockl = 4 and n TMBIM6 = 5). All bars represent mean ± SEM. In all tests, two-way ANOVA followed by Tukey’s multiple comparison test was performed. Statistical significance (p < 0.05) between samples is indicated in the figures as follows: *=p < 0.05; **=p < 0.01; ***=p < 0.001; ****=p < 0.0001.

Article Snippet: Using data on TMBIM6 mRNA expression levels from The Human Protein Atlas database (THPA) [ ], we confirmed through in silico analysis that TMBIM6 is expressed in various cortical regions of the human brain, with notably high expression in the midbrain and retina, two areas rich in DAergic cells [ , ] (Fig. ).

Techniques: In Vivo, Transduction, Expressing, Injection, Comparison