Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Box plots show TGM2 expression in 179 PDAC tissues and 171 corresponding non-tumor PDAC tissues. *** p < 0.001. B Kaplan–Meier curve analysis of the prognostic value of TGM2. C RT-qPCR and Western blot analysis of TGM2 expression in TT and TP tissues from PDAC patients. **** p < 0.0001 vs. TP. D RT-qPCR and Western blot analysis of TGM2 expression in normal pancreatic duct cells (HPNE) and PDAC cells (MIA-PACA-2, BxPC-3, PATU-8988S, PANC-1). **** p < 0.0001 vs. HPNE. E CCK-8 assay was performed to compare cell viability against PDAC and Gem-R PDAC cells. F RT-qPCR and Western blot analysis of TGM2 expression in PDAC and Gem-R PDAC cells. **** p < 0.0001 vs. PDAC cells. G The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. H Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in Gem-R PDAC cells. L The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. I The levels of TGM2 in the supernatants of PDAC and Gem-R PDAC cells. J RT-qPCR and Western blot analysis of TGM2 expression in TT and TP tissues from PDAC patients with Gem-R. K The levels of ATP and NH 4 + were detected in tumor and para-tumor tissues from PDAC patients with Gem-R. L The levels of TGM2 were examined in the supernatants of TT and TP tissues from PDAC patients with Gem-R. ** p < 0.01, **** p < 0.0001 vs. Gem-R TP. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A RT-qPCR and Western blot analysis of TGM2 expression in Gem-R PDAC cells. B Clone formation assay to detect cell proliferation. Scale bar: 100 μm. C . CCK-8 assay to detect cell viability. D Flow cytometry to detect cell apoptosis. E , F Transwell assay to detect cell invasion and migration. Scale bar: 100 μm. G Western blot analysis of GLS, LC3 Ⅱ/Ⅰ, and p62 expression in Gem-R PDAC cells. H The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. I The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. J The level of glutamate was detected in Gem-R PDAC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. sh-NC. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. sh-TGM2. &&& p < 0.001, &&&& p < 0.0001 vs. Glu. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Growth curves of tumors in nude mice. B Images of tumors and diagrams of tumor weight. C IHC assay to detect TGM2 expression in tumor tissues. Scale bar: 100 μm and 25 μm. D Western blot analysis of GLS, LC3 Ⅱ/Ⅰ, and p62 expression in tumor tissues. E The levels of ATP and NH 4 + were detected in tumor tissues. F The levels of 2-DG6P and lactate were detected in tumor tissues. G The level of glutamate was detected in tumor tissues. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. sh-NC. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. sh-TGM2. &&& p < 0.001, &&&& p < 0.0001 vs. Glu. n = 5.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Expressing, Western Blot

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A PCA analysis showed differences between groups. B TPM curves showed sequencing stability. C Venn diagram showing intersection of mitophagy pathway-related DEGs in different combinations of intergroup comparisons (sh-NC vs. Sh-TGM2, sh-TGM2 vs. sh-TGM2+Glu, Glu vs. sh-TGM2, sh-NC vs. Glu). D KEGG and GO analysis. E The expression analysis of P2RX7, AVPR1A, ADORA2A, and KMO by RNA sequencing. F RT-qPCR and Western blot analysis of P2RX7, AVPR1A, ADORA2A, and KMO expression in Gem-R PDAC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. sh-NC. ## p < 0.01, #### p < 0.0001 vs. sh-TGM2. &&& p < 0.001, &&&& p < 0.0001 vs. Glu. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Sequencing, Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A , B RT-qPCR and Western blot analysis of P2RX7 expression in PDAC and para-tumor tissues from PDAC patients with Gem-R. **** p < 0.0001 vs. Gem-R TP. C Western blot analysis of P2RX7 expression in Gem-R PDAC cells. ** p < 0.01 vs. sh-NC, #### p < 0.0001 vs. oe-NC. D Western blot analysis of P2RX7 and TGM2 expression in Gem-R PDAC cells. E Clone formation assay to detect cell proliferation. Scale bar: 100 μm. F CCK-8 assay to detect cell viability. G Flow cytometry to detect cell apoptosis. H , I Transwell assay to detect cell invasion and migration. Scale bar: 100 μm. J Representative TEM images of an autophagosome (arrow) in Gem-R PDAC cells. Scale bar: 2 μm. K Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in Gem-R PDAC cells. L The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. M The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. N The level of glutamate was detected in Gem-R PDAC cells. O The levels of TGM2 in the supernatants of Gem-R PDAC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. oe-NC. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. oe-P2RX7. & p < 0.05, && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 vs. Glu. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A . Western blot analysis of P2RX7 expression in Gem-R PDAC cells. B Clone formation assay to detect cell proliferation. Scale bar: 100 μm. C CCK-8 assay to detect cell viability. D Flow cytometry to detect cell apoptosis. E , G Transwell assay to detect cell invasion. Scale bar: 100 μm. F , H Transwell assay to detect cell migration. Scale bar: 100 μm. I Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in Gem-R PDAC cells. J The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. K The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. L The level of glutamate was detected in Gem-R PDAC cells. M The levels of TGM2 in the supernatants of Gem-R PDAC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. sh-NC. # p < 0.05, ### p < 0.001, #### p < 0.0001 vs. sh- P2RX7. & p < 0.05, && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 vs. Glu. @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, @@@@ p < 0.0001 vs. Glu+sh-P2RX7. $ p < 0.05, $$ p < 0.01, $$$ p < 0.001, $$$$ p < 0.0001 vs. Glu+ATP. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Western Blot, Expressing, Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Western blot analysis of P2RX7 expression in Gem-R PDAC cells. B Clone formation assay to detect cell proliferation. Scale bar: 100 μm. C CCK-8 assay to detect cell viability. D Flow cytometry to detect cell apoptosis. E , F Transwell assay to detect cell invasion and migration. Scale bar: 100 μm. G Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in Gem-R PDAC cells. H The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. I The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. J The level of glutamate was detected in Gem-R PDAC cells. K The levels of TGM2 in the supernatants of Gem-R PDAC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NC. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. oe-TGM2. & p < 0.05, && p < 0.01, &&& p < 0.001, & &&& p < 0.0001 vs. Glu. @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, @@@@ p < 0.0001 vs. Glu+oe-TGM2. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Western Blot, Expressing, Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Western blot analysis of TGM2 and P2RX7 expression in Gem-R PDAC cells. B Clone formation assay to detect cell proliferation. Scale bar: 100 μm. C CCK-8 assay to detect cell viability. D Flow cytometry to detect cell apoptosis. E , F . Transwell assay to detect cell invasion and migration. Scale bar: 100 μm. G Representative TEM images of an autophagosome (arrow) in Gem-R PDAC cells. Scale bar: 2 μm. H Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in Gem-R PDAC cells. I The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. J The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. K The level of glutamate was detected in Gem-R PDAC cells. L The levels of TGM2 in the supernatants of Gem-R PDAC cells. **** p < 0.0001 vs. Control. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. Cys. & p < 0.05, && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 vs. Cys+Glu. @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, @@@@ p < 0.0001 vs. Cys+oe-P2RX7. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Western Blot, Expressing, Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration, Control

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Growth curves of tumors in nude mice. B Images of tumors and diagrams of tumor weight. C Western blot analysis of parkin, LC3 Ⅱ/Ⅰ, p62, and GLS expression in tumor tissues. D The levels of ATP and NH 4 + were detected in tumor tissues. E The levels of 2-DG6P and lactate were detected in tumor tissues. F The level of glutamate was detected in tumor tissues. G The levels of TGM2 were examined in the supernatants of tumor tissues. **** p < 0.0001 vs. Control. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. Cys. & p < 0.05, && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 vs. Cys+Glu. @ p < 0.05, @@ p < 0.01, @@@ p < 0.001, @@@@ p < 0.0001 vs. Cys+oe-P2RX7. n = 5.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Western Blot, Expressing, Control

Journal: Cell Death Discovery

Article Title: TGM2-P2RX7 loop promotes gemcitabine resistance in pancreatic cancer by modulating glutamine metabolism and mitophagy

doi: 10.1038/s41420-025-02922-x

Figure Lengend Snippet: A Clone formation assay to detect cell proliferation. Scale bar: 100 μm. B CCK-8 assay to detect cell viability. C Flow cytometry to detect cell apoptosis. D , E Transwell assay to detect cell invasion and migration. Scale bar: 100 μm. F The levels of ATP and NH 4 + were detected in Gem-R PDAC cells. G The levels of 2-DG6P and lactate were detected in Gem-R PDAC cells. H The level of glutamate was detected in Gem-R PDAC cells. I The levels of TGM2 in the supernatants of Gem-R PDAC cells. *** p < 0.001, **** p < 0.0001 vs. Control. ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. Cys. & p < 0.05, && p < 0.01, &&&& p < 0.0001 vs. Cys+Glu+oe-P2RX7. n = 3.

Article Snippet: In accordance with the manufacturer’s instructions, we utilized the ATP Assay Kit (A095-1-1, Nanjing Jiancheng Bioengineering Institute), the EnzyChromTM Ammonia/Ammonium Assay Kit (ENH3-100, BioAssay Systems), the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma), the Lactate Assay Kit (A019-2-1, Nanjing Jiancheng Bioengineering Institute), the Glutamate Assay Kit (A074-1-1, Nanjing Jiancheng Bioengineering Institute), and the TGM2 ELISA Kit (CSB-E11797h, Cusabio) to detect the levels of ATP, NH 4 + , glucose uptake, lactate, glutamate, and TGM2, respectively.

Techniques: Tube Formation Assay, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration, Control