Journal: JID Innovations

Article Title: Peroxisomal Fatty Acid Oxidation and Glycolysis Are Triggered in Mouse Models of Lesional Atopic Dermatitis

doi: 10.1016/j.xjidi.2021.100033

Figure Lengend Snippet: Anaerobic glycolysis but not mitochondrial function is triggered in ft/ft mouse epidermis. ( a ) Lactate production and ( b ) relative mRNA level of Ldha in mouse epidermis (n = 5–9). ( c ) Intracellular levels of the intermediates of the Krebs cycle in mouse epidermis quantified by LC‒MS. Fold changes between the mean values of five mice per group are shown. The list of all quantified metabolites as well as respective fold changes are provided in Table 4 . ( d ) Relative mRNA expression of Mpc1 and Mpc2 in the epidermis of CTRL and ft/ft mice (n = 5–9). ( e ) Relative mRNA level of Pdk1 in mouse epidermis (n = 9–10). ( f ) Activity of Cx I and of Cx II of the respiratory chain in mouse epidermal cells (n = 9–10). Relative mRNA level of ( g ) Acadvl and ( h ) Acad9 in the epidermis of mice (n = 9–10). ( i ) Activity of mitochondrial ACADL measured in epidermal cells of CTRL and ft/ft mice (n = 10). ( j ) Relative mRNA level of Cpt1a in mouse epidermis (n = 5–9). Data were analyzed with a Student’s t -test. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. CTRL, control; Cx, complex; LC‒MS, liquid chromatography‒mass spectrometry; min, minute.

Article Snippet: muMpc1 , Mouse Mitochondrial pyruvate carrier 1 , Mm01316203_g1.

Techniques: Expressing, Activity Assay, Control

Journal: Scientific reports

Article Title: Effects of glucose metabolism pathways on nuclear and cytoplasmic maturation of pig oocytes.

doi: 10.1038/s41598-020-59709-6

Figure Lengend Snippet: Figure 4. Effects of down regulating related genes in DOs or CCM on in vitro maturation of pig oocytes. Graph (A) shows percentages of MII oocytes after DOs were cultured for 48 h in the NCSU-23 medium containing 0, 10 or 15 mM pyruvate (Pyr) with 10, 25 or 50 µM 4-CIN (CIN) or with 0.01, 0.03 or 0.05 µM rotenone (Rot). Graph B shows percentages of MII oocytes after DOs were cultured for 48 h in the NCSU-23 medium containing 0, 5 or 10 mM lactate (Lac) with 10, 25 or 50 mM sodium oxamate (Oxm) or with 10, 20 or 50 µM 4-CIN. Graphs (C–E) show MII percentages of cocultured DOs (Left Y axis) and levels of MPC1, NDUFV1 and LDHB (Right Y axis) after transfection of CCM with negative control (NC) siRNA, MPC1 siRNA (MP1, 2 or 3), NDUFV1 siRNA (ND1, 2 or 3), or LDHB siRNA (LD1, 2 or 3), respectively. Graphs (F,G) show MII percentages of DOs after microinjection with NC, MP3, ND1 or LD2 siRNAs. While DOs and the CCM transfected/injected with MPC1 or NDUFV1 siRNAs were cultured in NCSU-23 containing 15 mM pyruvate, DOs and the CCM transfected/injected with LDHB siRNA were cultured in NCSU-23 containing 10 mM lactate. In oocyte maturation experiments, each treatment was repeated 4 times with each replicate containing about 20 oocytes. Graphs (H,I) show ATP concentrations (ng/ml) in medium conditioned with CCM in NCSU-23 medium containing 5.6 mM glucose and 15 mM pyruvate, respectively, after transfection with G6PD siRNA-3 (G6-3), GAPDH siRNA-3 (GA-3) or MPC1 siRNA-3 (MP3). Each treatment was repeated 3 times with each replicate containing medium recovered from one culture well on different experimental days. a–f: Values with a different letter above bars differ significantly (P < 0.05).

Article Snippet: The primary antibodies used included rabbit anti-G6PD monoclonal antibodies (1:1000, ab993, Abcam Co., Ltd, Beijing, China), mouse anti-GAPDH monoclonal antibodies (1:1000, CW0100A, CWBio Co., Ltd, Beijing, China), rabbit anti-LDHB polyclonal antibodies (1:1000, 14824-1-AP, Proteintech Co., Ltd, Wuhan, China), rabbit anti-MPC1 polyclonal antibodies (1:200, TA315496S, ORIGENE Co., Ltd, Beijing, China), rabbit anti-NDUFV1 polyclonal antibodies (1:500, 11238-1-AP, Proteintech Co., Ltd, Wuhan, China), mouse anti-β-tubulin monoclonal antibodies (1:1000, 05–661, Merck Millipore), and mouse anti-β-actin monoclonal antibodies (1:1000, CW00096M, CWBio Co., Ltd).

Techniques: In Vitro, Cell Culture, Transfection, Negative Control, Microinjection, Injection

Journal: EMBO Reports

Article Title: Pyruvate metabolism guides definitive lineage specification during hematopoietic emergence

doi: 10.15252/embr.202154384

Figure Lengend Snippet: A FACS‐sorted HE, EHT, or HSC‐like cells were subcultured with or without UK5099 (10 µM). Subculture day 3 CD43 + /GPA + cell frequencies ± SEM relative to the controls for all populations are presented (HE, n = 5 biological replicates; EHT, n = 6 biological replicates; HSC‐like, n = 4 biological replicates; paired t ‐tests). B FACS‐sorted HE cells were subcultured with or without 1‐AA (4 mM). Subculture day 3 CD43 + GPA + cell frequencies ± SEM relative to the control are shown ( n = 4 biological replicates, paired t ‐test). C Fold change of expression of MPC1 and MPC2 ± SEM relative to HPRT1 in shRNA‐transduced cells compared to shScrambled (shScr) is shown ( n = 3 biological replicates, unpaired t ‐tests). Untr, untransduced. D HE cells were transduced with shScrambled (shScr), shMPC1, shMPC2, or both the day after the sort and day 3 CD43 + /GPA + cell frequencies ± SEM relative to shScr are presented ( n = 4 biological replicates; one‐way ANOVA test). Untr, untransduced. E FACS‐sorted HE cells were stained with CTV and fluorescence was assessed by flow cytometry for GPA + cells at day 3 of subculture with or without UK5099 (10 µM). Representative of n = 3. F, G FACS‐sorted HE, EHT, or HSC‐like cells were subcultured with or without UK5099 (10 µM). Subculture day 6 CD43 + (F) and CD43 + CD45 + (G) cell frequencies ± SEM relative to the control for all populations are presented (HE, n = 7 biological replicates; EHT, n = 7 biological replicates; HSC‐like, n = 4 biological replicates; paired t ‐tests). H FACS‐sorted HE cells were subcultured with or without 1‐AA (4 mM). Subculture day 6 CD43 + and CD43 + CD45 + cell frequencies ± SEM relative to the control are shown ( n = 3 biological replicates, paired t ‐test). I, J FACS‐sorted HE cells were subcultured for 3 days with or without UK5099 (10 µM). CTV for HE‐derived CD45 + cells (I) and HE‐derived HSC‐like cell frequencies ± SEM relative to the control ( n = 7 biological replicates, paired t ‐test) (J) are shown. K–N FACS‐sorted HE and EHT cells were subcultured with or without DCA (3 mM). Subculture day 3 (K, n = 3 biological replicates) and day 6 (L, HE, n = 5 biological replicates; EHT, n = 4 biological replicates) CD43 + GPA + cell frequencies ± SEM relative to the controls are shown (paired t ‐test). (M) CTV for HE‐derived GPA + cells at subculture day 3 is shown. Representative of n = 3. (N) Subculture day 6 CD43 + CD45 + cell frequencies ± SEM relative to the controls for both populations are shown (HE, n = 5 biological replicates; EHT, n = 4 biological replicates; paired t ‐tests). O Fold change of expression of PDK1 , PDK2 , PDK3 , and PDK4 ± SEM relative to HPRT1 in shRNA‐transduced cells compared to shScrambled (shScr) is shown ( n = 3 biological replicates, paired t ‐tests). P HE cells were transduced with shScrambled (shScr) or shPDK1, shPDK2, shPDK3, or shPDK4 the day after the sort and day 6 CD43 + /CD45 + cell frequencies ± SEM relative to shScr are presented ( n = 3 biological replicates; one‐way ANOVA tests). Q CTV for HE‐derived CD45 + cells are shown. Representative of n = 3. R HE‐derived HSC‐like cell frequencies ± SEM relative to the control ( n = 4 biological replicates, paired t ‐test) are shown. S EdU incorporation ± SEM into HE cells was assessed by flow cytometry after a 24‐h pulse at days 1 and 2 of subculture with or without UK5099 (10 µM) or DCA (3 mM) ( n = 3 biological replicates). T, U Percentages of CFU assay colony types ± SEM obtained from HE cells subcultured with the indicated compounds for 3 days (U) ( n = 3 biological replicates, two‐way ANOVA test) or 6 days (T) ( n = 5 biological replicates, two‐way ANOVA test). CFU, colony‐forming unit; E, erythroid; M, macrophage; G, granulocyte; GEMM, mixed. V EryD and EryP CFU‐Es obtained from HE cells subcultured with the indicated compounds for 3 days. Scale bars, 100 µm. W Fold change in the expression of HBA1‐2 transcripts ± SEM normalized to KLF1 in CFUs obtained from HE cells treated with UK5099 (10 µM) or DCA (3 mM) relative to non‐treated cells ( n = 3 biological replicates, paired t ‐test). X Plots showing percentages of CD45 + CD56 + cells obtained following 35‐day co‐culture of 3‐day subcultured HE cells with OP9‐DL1 stroma. During the 3‐day subculture, HE cells were treated with the indicated compounds. Data information: ns, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The taqman probes used for this assay are MPC1 (Hs00211484_m1), MPC2 (Hs00967250_m1), PDK1 (Hs01561847_m1), PDK2 (Hs00176865_m1), PDK3 (Hs00178440_m1), PDK4 (Hs01037712_m1), LSD1/KDM1A (Hs01002741_m1), and HPRT1 (Hs02800695_m1).

Techniques: Control, Expressing, shRNA, Transduction, Staining, Fluorescence, Flow Cytometry, Derivative Assay, Colony-forming Unit Assay, Co-Culture Assay

Journal: EMBO Reports

Article Title: Pyruvate metabolism guides definitive lineage specification during hematopoietic emergence

doi: 10.15252/embr.202154384

Figure Lengend Snippet:

Article Snippet: The taqman probes used for this assay are MPC1 (Hs00211484_m1), MPC2 (Hs00967250_m1), PDK1 (Hs01561847_m1), PDK2 (Hs00176865_m1), PDK3 (Hs00178440_m1), PDK4 (Hs01037712_m1), LSD1/KDM1A (Hs01002741_m1), and HPRT1 (Hs02800695_m1).

Techniques: Recombinant, Saline, Flow Cytometry, Gene Expression, RNA Sequencing, Software

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a Western blotting showing the expression of MPC1 in nine pairs LAC tumor tissue (T) and adjacent normal tissues (N). b Quantitative analysis of MPC1 protein expression in nine pairs LAC tumor tissue and adjacent normal tissues. c Expression of MPC1 mRNA in 15 pairs of LAC tumor tissues and adjacent normal tissues. d qRT-PCR and western blotting showing the expression of MPC1 in LAC cells (A549, H1299, H1975) and normal human bronchial epithelial cells (HBE). e Representative IHC image for LAC patients with MPC1 low and MPC1 high , Scare bar = 50 μm. f Kaplan–Meier analysis of overall survival rate (OS) in LAC patients with MPC1 low and MPC1 high . g Kaplan–Meier analyses of MPC1 in LAC patients from GEO database ( n = 1926). Data are expressed as the mean ± SD,* P <0.05, *** P <0.001

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Western Blot, Expressing, Quantitative RT-PCR

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: Univariate and multivariate analysis for overall survival in LAC patients

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Significance Assay, Expressing

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a , b The representative images ( a ) and numbers ( b ) of tumorsphere in OE MPC1 cells and Ctrl cells by tumorsphere formation assay, scale bar = 100 µm. c , d The representative images ( c ) and numbers ( d ) of tumorsphere in si MPC1 and siNC LAC cells, and cells treated with DMSO or UK5099 (40 μM) by tumorsphere formation assay, scale bar = 100 µm. e , f Western blotting analysis of the abundances of NANOG, OCT4, and SOX2 in up- ( e ) and downregulated ( f ) MPC1 groups as compared with control groups. Data are expressed as the mean ± SD, ** P <0.01

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Tube Formation Assay, Western Blot, Control

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a , b The representative images ( a ) and numbers ( b ) of motile OE MPC1 cells and Ctrl cells in the transwell invasion and migration assay (scale bar = 50 µm). c , d The representative images ( c ) and numbers ( d ) of motile siNC, si MPC1 in H1299 cells; DMSO- and UK5099-H1299 cells in the transwell invasion and migration assay (scale bar = 50 µm). e , f Immunofluorescence showing the representative images ( e ) and numbers ( f ) of invadopodia with OE MPC1 or Ctrl, the nuclei were stained with DAPI (blue) (Scale bar = 20 µm). g , h Immunofluorescence showing the representative images ( g ) and numbers ( h ) of invadopodia with si MPC1 or siNC, the nuclei were stained with DAPI (blue) (scale bar = 20 µm), Data are expressed as the mean ± SD, * P <0.05, ** P <0.01, *** P <0.001

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Migration, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a Identification of STAT3 from the intersection between MPC1 interacting protein set by MS and the KEGG pathway in cancer gene set from GSEA. b , c Co-IP analysis of STAT3 interaction with MPC1 in A549 and H1299 MPC1-overpxpression cells. d The mitochondria was isolated from the cytoplasm, and Co-IP analysis showing that STAT3 interacts with MPC1 in the mitochondria in A549 and H1299 MPC1-overpxpression cells. e Western blotting showing that the protein expression levels of STAT3, P-STAT3(S727), P-STAT3(Y705), and MPC1 in A549 and H1299 cell with stable infection of OE MPC1 or Ctrl vector. f The mitochondria was isolated from the cytoplasm, mitochondria (mito) and cytoplasm without mitochondria (cyto) were prepared, western blotting showing that the protein expression levels of P-STAT3(Y705), STAT3, MPC1, and mitochondrial reference COX IV in A549 and H1299 cell with stable infection of OE MPC1 or Ctrl vector. g The nuclear and cytoplasmic extracts were prepared. Western blotting showing that overexpression of MPC1 decreased the expression of P-STAT3(Y705) in the nuclei. h IHC showing the expression of P-STAT3(Y705) in the MPC1 low LAC tissues ( n = 11) and MPC1 high ( n = 9) LAC tissues, Scale bar = 50 μm. i IHC score of P-STAT3(Y705) in the MPC1 low LAC tissues ( n = 11) and MPC1 high LAC tissues ( n = 9). Data are expressed as the mean ± SD, * P <0.05

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Co-Immunoprecipitation Assay, Isolation, Western Blot, Expressing, Infection, Plasmid Preparation, Over Expression

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a Western blotting showing the protein expression levels of STAT3 and P-STAT3(Y705) in A549 cells treated with IL-6 for 24 h at the indicated dose. b , c The representative images ( b ) and numbers ( c ) of tumorspheres from the Ctrl and OE MPC1 A549 and H1299 cells treated with or without IL-6 (100 ng/ml). Scale bar = 100 µm. d The number of motile Ctrl and OE MPC1 A549 and H1299 cells treated with or without IL-6 (100 ng/ml). e Western blotting showing the expression of SOX2, MMP2, and MPC1 in Ctrl or OE MPC1 A549 and H1299 cells treated with or without IL-6 (100 ng/ml). Data are expressed as the mean ± SD, * P <0.05, ** P <0.01

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: MPC1 deficiency accelerates lung adenocarcinoma progression through the STAT3 pathway

doi: 10.1038/s41419-019-1324-8

Figure Lengend Snippet: a Nude mice were injected subcutaneously in the right flanks with Ctrl or MPC1-overexpression A549 cells. After 28 days, xenografts were harvested and photographed. b , c Quantitative analysis of the volume ( b ) and weight ( c ) of the xenograft tumors. d IHC staining showing the expression of MPC1, P-STAT3(Y705), SOX2, and MMP2 in the xenograft tumors derived from OE MPC1 and Ctrl cells, Scale bar = 25 µm. e Schematic diagram of the regulatory mechanism of MPC1 mediates STAT3 dephosphorylation and inhibits the stemness and invasion of LAC cells. Data are expressed as the mean ± SD,* P <0.05, ** P <0.01, *** P <0.001

Article Snippet: The siRNAs were list as follows, MPC1 siRNAs were custom synthesized from GenePharma (Shanghai, China) and were pooled as (5′-UCCCTCTUTTUCTATTCTTTU-3′) and (5′-UUCTTATCAAACACUAUATUA-3′).

Techniques: Injection, Over Expression, Immunohistochemistry, Expressing, Derivative Assay, De-Phosphorylation Assay

Journal: The EMBO Journal

Article Title: The yeast mitochondrial pyruvate carrier is a hetero‐dimer in its functional state

doi: 10.15252/embj.2018100785

Figure Lengend Snippet: The individual histidine‐tagged Mpc1 (Mpc1his), histidine‐tagged Mpc2 (Mpc2his) or the Mpc1/Mpc2 hetero‐complex (Mpc1/Mpc2his) was expressed in the triple mpc knock‐out strain SHY15 and detected with antibodies raised against Mpc1 (left) or against the histidine tag (right). SDS–PAGE analysis of purified Mpc1, Mpc2 and Mpc1/Mpc2 proteins, visualised with Coomassie Blue stain. As the yield of the purified Mpc2 was very low, the protein was not visible with Coomassie Blue stain, but was identified on the gel by peptide mass finger printing ( Table EV1 ). The stability of the purified Mpc1/Mpc2 or Mpc2 was assessed via the CPM method. Thermal denaturation profiles ( upper panel ) were used to calculate the first derivative of the data ( lower panel ). The number is the apparent melting temperature for the Mpc1/Mpc2 hetero‐complex. Time course of pyruvate homo‐exchange by the Mpc1/Mpc2 hetero‐complex in liposomes in comparison with empty liposomes at a ΔpH of 1.6 ( n = 2).

Article Snippet: The codon‐optimised gene sequences for Mpc1 (UniProt: P53157), Mpc2 (UniProt: P38852) and Mpc3 (UniProt: P53311) from S. cerevisiae were synthesised (GenScript) and cloned into the bidirectional expression vector pBEVY‐GU [gift from Charles Miller; Addgene plasmid # 51229 (Miller et al , )].

Techniques: Knock-Out, SDS Page, Purification, Staining, Liposomes, Comparison

Journal: Nature Communications

Article Title: Metabolic imaging across scales reveals distinct prostate cancer phenotypes

doi: 10.1038/s41467-024-50362-5

Figure Lengend Snippet: a Schematic representation of the proposed differences in the metabolic fate of [1- 13 C]pyruvate between the benign and malignant prostatic epithelium, with the latter showing increased mitochondrial pyruvate uptake via AR-regulated MPCs to fuel both the restored TCA cycle and FASN-catalysed fatty acid biosynthesis. b Representative fluorescent RNAscope images of epithelial mRNA LDHA (gold) and LDHB (white) expression, along with IHC images of epithelial MPC2, FASN, and AR expression in the benign and malignant glands. c Scatterplots with bars comparing the expression of epithelial LDH, MPC2, FASN, and AR density between the benign ( n = 11 samples) and tumour ( n = 11 samples) areas from the HP- 13 C-MRI cohort ( n = 7 patients). d Top: Plots comparing the log-transformed epithelial and stromal MPC1 and MPC2 densities in the benign ( n = 11 samples) and tumour ( n = 11 samples) areas from the HP- 13 C-MRI cohort ( n = 7 patients). Bottom: Spearman’s correlation plots comparing tumour epithelial MPC2 density against tumour epithelial FASN and AR densities, as well as HP- 13 C-MRI-derived [1- 13 C]lactate SNR. e Mixed box-and-whisker and scatterplots comparing TCGA-PRAD derived bulk mRNA expression of LDHA , LDHB , total LDH, MPC2 , FASN , and AR between benign ( n = 45) and tumour ( n = 293) prostatectomy samples from n = 293 patients. f Average expression dot plots comparing single-cell RNA-seq epithelial expression of the same genes from publicly available EGAS00001005787 (left) and GSE176031 (right) datasets. g H&E map of a surgical specimen including areas of benign and malignant prostate with corresponding spatial transcriptomics maps demonstrating the expression of total LDH, MPC2 , and FASN obtained from a publicly available EGAS00001006124 dataset. In c – e , lines are median values and bars are interquartile ranges, with P derived using the two-sided Mann–Whitney U test or Wilcoxon signed-rank test, as appropriate. Scale bars denote 5–10 μm. Source data are provided as a Source Data file. d was created with BioRender.com and released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

Article Snippet: To assess biological factors such as epithelial capacity for [1- 13 C]pyruvate uptake, tissue vascularity, and inferred [1- 13 C]pyruvate metabolic fate, we used IHC to stain the HP- 13 C-MRI cohort samples for the following targets: MCT1 (membrane pyruvate importer; Cat. No. HPA003324, Atlas Antibodies, Bromma, Sweden), MPC1 and MPC2 (mitochondrial pyruvate carriers; Cat. No. PAB28306, Abnova, Taipei, Taiwan for MPC1; Cat. No. D417G, Cell Signalling Technology, Danvers MA, USA for MPC2), FASN (cytosolic fatty acid synthase; Cat. No. 3180, Cell Signalling Technology, Danvers MA, USA), AR (nuclear receptor regulating the expression of MPCs and FASN; Cat. No. NCL-AR-318, Novocastra, Newcastle, UK), HIF-1α (nuclear transcription factor regulating the expression of LDHA ; Cat. No. ab51608, Abcam, Cambridge, UK), and CD31 (endothelial biomarker; Cat. No. M0823, Dako, Santa Clara CA, USA).

Techniques: RNAscope, Expressing, Transformation Assay, Derivative Assay, Whisker Assay, RNA Sequencing, MANN-WHITNEY

Journal: Nature Communications

Article Title: Metabolic imaging across scales reveals distinct prostate cancer phenotypes

doi: 10.1038/s41467-024-50362-5

Figure Lengend Snippet: a , b Whole-mount H&E, ADC, along with [1- 13 C]pyruvate and [1- 13 C]lactate SNR maps demonstrating the presence of large, cellular ISUP GG3 lesions with dominant ICC component that were HP- 13 C-MRI occult compared to contralateral small-volume foci of ISUP GG2 disease with <5% non-cribriform GP4 glands. c Comparator case of a HP- 13 C-MRI-visible large cribriform ISUP GG3 tumour that was also visible on 1 H-MRI ADC. d Representative H&E slides, along with corresponding IHC-derived CD31, MCT1, MPC2, FASN, AR, and RNAscope-derived total LDH images obtained from HP- 13 C-MRI-visible and HP- 13 C-MRI-occult lesions shown in a – c . e Mixed box-and-whisker and scatterplots comparing [1- 13 C]pyruvate and [1- 13 C]lactate SNR, epithelial cell density, CD31 density, as well as epithelial MCT1, LDH, MPC2, FASN, and AR density between HP- 13 C-MRI-visible ( n = 13 samples) and HP- 13 C-MRI-occult ( n = 2 samples) lesions from the hyperpolarised MRI cohort ( n = 8 patients). a – d include images from three separate patients; imaging and staining were not repeated. In e , the data points for the 13 C-visible ICC tumour are coloured in black. Scale bars in a – c and d denote 5 mm and 5–50 μm, respectively. Source data are provided as a Source Data file.

Article Snippet: To assess biological factors such as epithelial capacity for [1- 13 C]pyruvate uptake, tissue vascularity, and inferred [1- 13 C]pyruvate metabolic fate, we used IHC to stain the HP- 13 C-MRI cohort samples for the following targets: MCT1 (membrane pyruvate importer; Cat. No. HPA003324, Atlas Antibodies, Bromma, Sweden), MPC1 and MPC2 (mitochondrial pyruvate carriers; Cat. No. PAB28306, Abnova, Taipei, Taiwan for MPC1; Cat. No. D417G, Cell Signalling Technology, Danvers MA, USA for MPC2), FASN (cytosolic fatty acid synthase; Cat. No. 3180, Cell Signalling Technology, Danvers MA, USA), AR (nuclear receptor regulating the expression of MPCs and FASN; Cat. No. NCL-AR-318, Novocastra, Newcastle, UK), HIF-1α (nuclear transcription factor regulating the expression of LDHA ; Cat. No. ab51608, Abcam, Cambridge, UK), and CD31 (endothelial biomarker; Cat. No. M0823, Dako, Santa Clara CA, USA).

Techniques: Derivative Assay, RNAscope, Whisker Assay, Imaging, Staining

Journal: Nature Communications

Article Title: Metabolic imaging across scales reveals distinct prostate cancer phenotypes

doi: 10.1038/s41467-024-50362-5

Figure Lengend Snippet: a Scatterplots with bars comparing clinical [1- 13 C]lactate labelling between tumours divided based on the percentage (far left; n = 7 samples for low %GP4 and n = 8 samples for high %GP4 lesions, respectively) and histological subtype (second from left; n = 7 samples for low %GP4, n = 5 samples for non-cribriform high %GP4, and n = 3 samples for cribriform high %GP4 lesions, respectively) of GP4 disease from n = 8 patients. Intergroup comparisons of 1 H-MRI-derived tumour ADC and K trans , as well as tissue-based epithelial MCT1 density are also presented. b Mixed box-and-whisker and scatterplots comparing tissue-based total epithelial LDH, nuclear HIF-1α, MPC2, FASN, and nuclear AR between ROIs harbouring individual GP3 ( n = 50), non-cribriform GP4 ( n = 22), and GP4 ICC ( n = 17) glands from n = 8 patients. c Representative H&E, RNAscope, and IHC images illustrating differential expression of tissue-based metabolic biomarkers between the three Gleason pattern glands. In a , b , the data are presented as median (denoted by the bars or boxes) with interquartile range (denoted by the bars or whiskers). P were derived using the two-sided Mann–Whitney U test. Source data are provided as a Source Data file. Scale bars denote 5–50 μm.

Article Snippet: To assess biological factors such as epithelial capacity for [1- 13 C]pyruvate uptake, tissue vascularity, and inferred [1- 13 C]pyruvate metabolic fate, we used IHC to stain the HP- 13 C-MRI cohort samples for the following targets: MCT1 (membrane pyruvate importer; Cat. No. HPA003324, Atlas Antibodies, Bromma, Sweden), MPC1 and MPC2 (mitochondrial pyruvate carriers; Cat. No. PAB28306, Abnova, Taipei, Taiwan for MPC1; Cat. No. D417G, Cell Signalling Technology, Danvers MA, USA for MPC2), FASN (cytosolic fatty acid synthase; Cat. No. 3180, Cell Signalling Technology, Danvers MA, USA), AR (nuclear receptor regulating the expression of MPCs and FASN; Cat. No. NCL-AR-318, Novocastra, Newcastle, UK), HIF-1α (nuclear transcription factor regulating the expression of LDHA ; Cat. No. ab51608, Abcam, Cambridge, UK), and CD31 (endothelial biomarker; Cat. No. M0823, Dako, Santa Clara CA, USA).

Techniques: Derivative Assay, Whisker Assay, RNAscope, Quantitative Proteomics, MANN-WHITNEY