cd3 bio rad mca1477 synthetic peptide 12 sequence  (Bio-Rad)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: RGX-019-MMAE inhibits leukemia progression by targeting MER proto-oncogene tyrosine kinase (MERTK) in acute myeloid leukemia

doi: 10.1186/s13046-026-03657-y

Figure Lengend Snippet: The antibody-drug conjugate RGX-019-MMAE induced cytotoxic activity in MERTK-expressing AML cell lines. A MERTK expression in the indicated AML cell lines; leukemic cells (1 × 10 6 ) were stained with anti-MERTK-APC antibody, and the expression was measured by flow cytometry. Data are plotted as mean values with error bars representing standard error. B - F Bar graph showing the percentage of relative luminescence in Kasumi 1 ( B ), OCI-AML3 ( C ), MOLM-13 ( D ), MOLM-14 ( E ), and MV4-11 ( F ) cells treated with the indicated concentrations of RGX-019-MMAE and monoclonal antibody RGX-019. Data are plotted as mean values with error bars representing standard error (Student unpaired t -test). * p ≤0.05, ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001. UT = untreated

Article Snippet: Mouse IgG1/kappa light chain MERTK-specific monoclonal antibodies were generated in mice by immunization with Fc-tagged human MERTK peptide (R&D Systems, Minneapolis, MN) USA).

Techniques: Activity Assay, Expressing, Staining, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: RGX-019-MMAE inhibits leukemia progression by targeting MER proto-oncogene tyrosine kinase (MERTK) in acute myeloid leukemia

doi: 10.1186/s13046-026-03657-y

Figure Lengend Snippet: The antibody-drug conjugate RGX-019-MMAE induced cytotoxic activity in MERTK-expressing AML primary cells. A - D Bar graphs show the percentage of relative luminescence in primary AML cells, RGX946 ( A ), RGX694 ( B ), RGX702 ( C ), and RGX470 ( D ) treated with the indicated concentrations of RGX-019-MMAE or the monoclonal antibody RGX-019. Data are plotted as mean values with error bars representing standard error (Student unpaired t -test). E Representative figures of colony-forming units for normal peripheral blood mononuclear cells in response to DMSO and indicated concentrations of RGX-019-MMAE and RGX-019. Data are plotted as mean values with error bars representing standard error (Student unpaired t -test) * p ≤0.05, ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001

Article Snippet: Mouse IgG1/kappa light chain MERTK-specific monoclonal antibodies were generated in mice by immunization with Fc-tagged human MERTK peptide (R&D Systems, Minneapolis, MN) USA).

Techniques: Activity Assay, Expressing

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) Schematic of the differentiation process with representative images of WT and MIC13 mut cells showing cellular morphology at different differentiation stages. Undifferentiated iPSC (at Day 0) display the typical oval or round shape, while differentiated induced hepatocytes (iHeps) (at Day 15) adopt a polygonal morphology, indicative of successful differentiation. Abbreviations: definitive endoderm (DE), hepatic endoderm (HE) and induced hepatocytes (iHeps). Scale bar: 100 μm (B) Quantitative PCR (qPCR) analysis was conducted at different differentiation stages using stage-specific gene markers: OCT4 (Octamer-binding transcription factor 4), SOX2 (Sex determining region Y-box 2), NANOG (Homeobox gene) for pluripotency in iPSCs; SOX17 (Sex determining region Y-box 17) and FOXA2 (Forkhead box 2) for definitive endoderm; HNFa (Hepatocyte Nuclear Factor a) and CEPBa (CCAAT Enhancer Binding Protein Alpha) for hepatic endoderm; and ALB (Albumin), A1AT (Alpha-1-antitrypsin) and AFP (Alpha-fetoprotein) for hepatocytes (iHeps). While there was a stage-specific decrease of pluripotency markers, iHeps markers increase steadily over the course of differentiation, peaking at iHeps stage. Expression of intermediate stage markers emerge at the corresponding stage of differentiation. This verifies the course of differentiation. Notably, all the markers (except OCT4, SOX2) are unaffected by the MIC13 deficiency across all stages of differentiation, demonstrating that MIC13 is not required for hepatic lineage acquisition. Data is showing log 10 fold change of WT and MIC13 mut individually normalized to the ΔC t -values of the iPSC stage. It is represented as bar graph with mean ± SEM (n = 3 - 7). Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05, ns = non-significant, p -value >0.05. HE marker CEPBa is exempt from statistical analysis at iHeps stage due to n = 2. (C) Functional assessment using indocyanine green (ICG) staining of iHeps shows uptake and release of the compound indicating intact metabolic activity in both WT and MIC13 mut iHeps. Scale bar: 100 μm. (D) Periodic acid-Schiff (PAS) staining demonstrates presence of glycogen storage in both WT and MIC13 mut iHeps. Scale bar: 100 μm.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Real-time Polymerase Chain Reaction, Binding Assay, Expressing, Marker, Functional Assay, Staining, Activity Assay

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) Western blot analysis shows marked depletion of the MIC10-subcomplex (MIC10, MIC26, MIC27 and MIC13) in MIC13 mut iHeps, while the MIC60-subcomplex (MIC60, MIC19 and MIC25) remains largely unaffected, except for MIC25. (B) Representative TEM images of WT and MIC13 mut iHeps reveal abnormal cristae ultrastructure, including stacked or ring-like structure of the cristae. Scale bars: 500nm. (C) Quantitative analysis of TEM images show that while the number of cristae per mitochondrial sections remains unchanged between WT and MIC13 mut iHeps, there is a significant reduction in number of CJs in MIC13 mut iHeps, indicating impaired cristae organization. Data is shown as separate violin plot with range, median and interquartile values indicated from approximately 135 sections from three independent (n = 3) experiments. Statistical analysis was performed using Student’s t test. ∗∗∗∗ p -value ≤0.0001, ns = non-significant, p -value >0.05. (D) Representative confocal images of WT and MIC13 mut iHeps immunostained for HSP60 (mitochondria, red) and counterstained with Hoechst (nuclei, blue) revealed extensive mitochondrial fragmentation in MIC13 mut iHeps. Yellow arrowhead indicate fragmented mitochondria and yellow lines traces tubular mitochondria. Scale bar: 10µm. (E) Quantification of mitochondrial network morphology, shown as a bar graph representing the percentage of cells displaying tubular, intermediate or fragmented mitochondria reveals a significant reduction in cells with tubular networks and a corresponding increase in cells with fragmented networks in MIC13 mut iHeps (around 50 cells per experiment, n = 3, mean ± SEM). Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05, ∗∗∗ p -value ≤0.001, ns = non-significant, p -value >0.05. (F) Representative confocal microscopy images of WT and MIC13 mut iHeps immunostained with anti-DNA (mitochondrial nucleoids and nuclei, green) and anti-HSP60 (mitochondria, red), reveal no detectable alterations in nucleoid distribution within mitochondria in MIC13 mut iHeps. White arrow heads indicate mtDNA nucleoids within mitochondria. Scale bar: 10µm. (G) Quantification of nucleoid count per cell shows no significant change between WT and MIC13 mut iHeps. Data is represented as a violin plot indicating the range, median and interquartile values, with data from around 90 cells from two independent experiments. Statistical analysis was performed using Student’s t test. ns = non-significant, p -value >0.05. (H) Quantitative PCR of genomic DNA shows no significant differences in the copy number of three mitochondrial genes, MT-ND1 (mitochondrially encoded NADH dehydrogenase 1), MT-RNR2 (mitochondrially encoded 16S RRNA), and MT-CYB (mitochondrially encoded cytochrome-b) in MIC13 mut iHeps. Data represented as bar graph (n = 3, mean ± SEM). Statistical analysis was performed using Student’s t test. ns = non-significant, p -value >0.05.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Western Blot, Confocal Microscopy, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) Heatmap of differentially regulated genes (DEGs) in WT and MIC13 mut iPSCs and iHeps reveals strong transcriptional differences between cell types, confirming robust differentiation, while MIC13-dependent changes within each cell type were subtler and largely cell-type specific. (B) Venn diagram illustrating overlaps of DEGs across group comparisons. The iHep-specific differences between WT and MIC13 mut (bold outline) include 1518 DEGs, with 600 upregulated and 918 downregulated genes (Bonferroni-corrected p value, Fold change ≥ 1.5). (C) GO biological process term enrichment analysis in MIC13 mut iHeps was performed on all significant DEGs (|log 2 Fold change| ≥ 0.59) as well as separately on upregulated (log 2 Fold change > 0.59) and downregulated (log 2 Fold change < −0.59). Analysis of all DEGs identifies pathways associated with extracellular matrix (ECM) organization, mitotic processes, cell migration and fatty acid metabolism. Upregulated genes are enriched for metabolic pathways, particularly lipid and amino acid metabolism, whereas downregulated genes are enriched for mitotic processes and ECM organization. (D) Proteomics analysis of MIC13 mut iHeps supports the transcriptomics trends. Among the top 30 upregulated proteins in MIC13 mut iHeps, GO-term based enrichment identifies ECM organization and coagulation as most enriched categories, whereas for the top 30 downregulated proteins, the highest categories correspond to lipid metabolic and protein catabolic processes. (E) STRING network analysis of significantly altered proteins in MIC13 mut iHeps reveals six distinct protein clusters, including Fibrinolysis, ECM proteoglycans, Fibrin clot formation, and Glutamine family amino acid catabolism, among others. Clusters are highlighted by distinct node colors.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Migration, Coagulation

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet:

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques:

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) ChemRICH Plot of significantly altered metabolites in MIC13 mut iHeps reveals strong enrichment of dipeptides and amino acids, indicating broad change in amino acid metabolism and protein turnover. (B) In MIC13 mut iHeps, multiple metabolic pathways are altered. TCA cycle intermediates, including α-ketoglutarate, fumarate and malate, are significantly increased, suggesting altered TCA cycle activity. Urea cycle related metabolites, including aspartate, argininosuccinate and arginine, are significantly increased in MIC13 mut iHeps, reflecting altered nitrogen metabolism. Intermediates of methionine cycle, including methionine, S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), and betaine were significant elevated in MIC13 mut iHeps, highlighting increased flux through methionine cycle and impaired methylation activity. Metabolites of the transsulfuration pathway, including cystathionine, hypotaurine, and taurine, are elevated, indicating enhanced flux towards glutathione biosynthesis and antioxidative capacity in response to redox stress in MIC13 mut iHeps. Data from five biological replicates. Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05, ∗∗ p -value ≤0.01, ∗∗∗ p -value ≤0.001, ∗∗∗∗ p -value ≤0.0001. (C) Schematic overview summarizing the metabolic rewiring of interconnected pathways in MIC13 mut iHeps. The diagram integrates metabolomic, transcriptomic and proteomic changes at key metabolic nodes, highlighting coordinated alterations across the TCA cycle, urea cycle, methionine cycle, and transsulfuration pathway. Elevated fumarate and aspartate link TCA and urea cycle flux, while homocysteine is diverted towards both remethylation and transsulfuration, reflecting simultaneous adjustments in methylation capacity and redox balance. Increased protein turnover provides amino acid that feed into the TCA cycle at multiple entry points. Additionally, the transcripts or proteins of key branch-point enzymes are altered, illustrating a coordinated transcriptional, proteomic and metabolic compensatory response to mitochondrial dysfunction. Abbreviations: ACO1 (Aconitase 1), IDH1 (Isocitrate dehydrogenase 1), ASL (Argininosuccinate lyase), ASS1 (Argininosuccinate synthase), BHMT1/2 (Betaine-homocysteine methyltransferase 1/2), CBS (Cystathionine B-synthase), ACAT1 (acetyl-CoA acetyltransferase 1), BDH2 (3-hydroxybutrate dehydrogenase 2). (D) Bile acid and its conjugates are increased, suggesting enhanced hepatic biosynthetic activity. (E) Lactate levels were elevated, indicating mitochondrial stress and increased glycolytic compensation. (F) Oxygen consumption rates (OCR) measured using Seahorse mitochondrial stress test reveals reduced mitochondrial respiration in MIC13 mut iHeps, consistent with widespread metabolic dysregulation and mitochondrial dysfunction. Representative traces from the complete experimental run is shown. (B, D, E) Data from five biological replicates. Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05, ∗∗ p -value ≤0.01, ∗∗∗ p -value ≤0.001, ∗∗∗∗ p -value ≤0.0001.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Activity Assay, Methylation, Metabolomic

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) In MIC13 mut iHeps, fatty acid metabolism is altered. Long-chain saturated and unsaturated acylcarnitines are significantly decreased, indicating impaired fatty acid activation and mitochondrial import. Conversely, short-chain acylcarnitines were increased, reflecting incomplete β-oxidation. Long-chain fatty acids, including palmitic and oleic acids, are elevated, consistent with impaired conversion to acylcarnitine. Ketone bodies are increased, suggesting a metabolic shift towards a compensatory response to altered TCA cycle flux and impaired fatty acid oxidation. Data from five biological replicates. Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05, ∗∗ p -value ≤0.01, ∗∗∗ p -value ≤0.001, ∗∗∗∗ p -value ≤0.0001. (B) Proteomics analysis shows significant downregulation of key proteins involved in fatty acid metabolism and lipid droplet biology, including CPT1A (Carnitine Palmitoyltransferase 1A), ACOT2 (Acyl-CoA Thioesterase 2), EHHADH (Enoyl-CoA Hydratase and 3-Hydroxyacyl CoA Dehydrogenase), CROT (Carnitine O-Octanoyltransferase) and PLIN2 (Perilipin 2), supporting broad defects in fatty acid activation, trafficking, oxidation and lipid storage. Data are represented as fold change relative to WT, error bars represent upper and lower limits calculated by back-transforming the log 2 fold change ± SD. Data is collected from four biological replicates. (C) STRING network of DEGs from the MIC13 mut iHeps GO BP term enrichment as grouped by function. Specific genes show coordinated dysregulation, thus providing evidence of systemic lipid metabolic defects in MIC13 mut iHeps. Node colors represent the log 2 fold changes. (D) BODIPY staining (green) of WT and MIC13 mut iHeps, counterstained with Hoechst (blue), shows marked reduction in lipid droplet accumulation in MIC13 mut iHeps. Arrow head marks lipid droplets. Scale bar: 10 μm. (E) Representative TEM images confirmed reduced lipid droplets in MIC13 mut iHeps. Lipid droplets were identified as round electron-lucent structures, marked by yellow arrow head. Scale bar: 1 μm. (F) Quantification of number of lipid droplets using TEM images demonstrates a significant reduction in lipid droplets abundance. Data are represented as violin plot where range, median and interquartile values are indicated. Data from around 80 sections from three independent experiments (n = 3). Statistical analysis was performed using Student’s t test. ∗∗∗∗ p -value ≤0.0001

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Activation Assay, Staining

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: (A) Heatmap of differentially regulated genes (DEGs) in MIC13 mut iHep by GO term-based enrichment for ECM-related genes. DEGs were considered statistically significant with a cut-off fold change of ±1.5 and Bonferroni correction p ≤ 0.05. (B) STRING network of DEGs from the MIC13 mut iHeps GO BP term enrichment as grouped by function (colored nodes). The groups show various functions within extracellular matrix organization. Log 2 fold changes are shown as a colored borders and edges of the respective nodes, revealing coordinative regulation (e.g. genes from the group “ECM disassembly” are mostly downregulated, whereas genes from “Elastic fiber formation” are upregulated. (C) Proteomics analysis of MIC13 mut iHeps shows that some of the most differentially altered proteins are ECM components, including COL1A1 (collagen type 1 alpha 1), FBLN2 (fibulin-2), FGG and FGB (fibrinogen gamma and beta chains), COL6A1 (collagen type V1 alpha 2), ITGB6 (integrin beta 6), Serine peptidase inhibitor, Kazal type 1 (SPINK1), TNC (tenasin C), and CCN1 (Cellular communication network factor 1). Data are represented as fold change relative to WT, error bars represent upper and lower limits calculated by back-transforming the log 2 fold change ± SD. Data collected from four biological replicates. Statistical analysis was performed using Student’s t test. ∗ p -value ≤0.05. (D) Immunofluorescence of COL1A1 (red) in iHeps with Hoechst nuclear counterstain (blue) reveals increased intracellular COL1A1 in MIC13 mut iHeps, suggesting either impaired secretion or biogenesis. Scale bar: 10μm. (E) Quantification of the COL1A1 signal shows a significant increase of COL1A1 abundance inside the MIC13 mut iHeps. Data is shown as violin plot with range, median and interquartile values indicated from approximately 100 cells analyzed from two independent experiments. Statistical analysis was performed using Student’s t test. ∗∗∗ p -value ≤0.001. (F) Wound healing assay shows accelerated migration of MIC13 mut iHeps, as determined by wound closure over time, reflecting altered cell-ECM interactions. Scale bar: 100 μm (G) Quantification of the wound closure area demonstrates reduction in the area in MIC13 mut iHeps, indicating faster movement. Data are represented as mean ± SEM from three independent experiments. Statistical analysis was performed using Student’s t test. ∗∗ p -value ≤0.01.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Immunofluorescence, Wound Healing Assay, Migration

Journal: bioRxiv

Article Title: MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

doi: 10.64898/2026.01.16.699229

Figure Lengend Snippet: Loss of MIC13 leads to mitochondrial cristae defects without directly affecting the ETC or mtDNA maintenance, leading to extensive metabolic rewiring in hepatocytes. These changes include increased protein turnover and amino acid utilization, altered TCA cycle activity, disruption in urea cycle, methionine cycle, transsulfuration pathways, and impaired redox homeostasis and enhanced bile acid synthesis. Lipid metabolism was altered characterized by reduced lipid droplets and enhanced ketogenesis, decrease long-chain acylcarnitines (AC), and increased short-chain acylcarnitines and fatty acids, reflecting impaired fatty acid activation and oxidation. Reduced CPT1A and other fatty acid metabolism enzymes of mitochondria and peroxisomes indicate defective lipid handling. At the cellular levels, these metabolic disturbances were associated with altered expression of transcriptional programs related to ECM organization, mitosis, and metabolic regulation, leading to ECM remodeling, intracellular collagen accumulation and enhanced cell migration. Together, this model provides mechanistic insight into how systemic MIC13 deficiency drives mitochondrial liver disease, by linking primary cristae defects to metabolic failure and downstream cellular ECM alterations.

Article Snippet: The following antibodies were used: MIC10 (Abcam, 84969), MIC13 (custom made by Pineda (Berlin) against human MIC13 peptide CKAREYSKEGWEYVKARTK) , MIC19 (Proteintech, 25625-1-AP), MIC25 (Proteintech, 20639-1-AP), MIC26 (Thermo fisher Scientific, MA5-15493), MIC27 (Sigma-Aldrich, HPA000612-100UL), MIC60 (Abcam, ab110329), HSP60 (sigma, SAB4501464).

Techniques: Activity Assay, Disruption, Activation Assay, Expressing, Migration