mpc1 sirna  (Shanghai GenePharma)

Journal: Redox Biology

Article Title: Hypobaric hypoxia-driven energy metabolism disturbance facilitates vascular endothelial dysfunction

doi: 10.1016/j.redox.2025.103675

Figure Lengend Snippet: Blocking MPC1 impairs vascular endothelial function though disrupting energy homeostasis. Diagram of cell energy metabolism after blocking MPC1 using siRNA or UK-5099 (10 μM) ( A ); ( B–H ) RAECs was treated with UK-5099 for 24 h to block MPC1. Representative images of immunofluorescence analysis of eNOS ( B ); The protein expression of eNOS, p-eNOS, ZO-1, Claudin-5, Occludin and VE-Cadherin was detected. ∗ p < 0.05 (n = 4) ( C ); The protein expression of MCT4 in RAECs. ∗ p < 0.05 (n = 4) ( D ); Lactate content in the supernatant of RAECs culture. ∗ p < 0.05 (n = 3) ( E ); Lactate content in the cytosol of RAECs, normalized using intracellular protein content. ∗ p < 0.05 (n = 3) ( F ); Representative images of the mitochondrial membrane potential of RAECs detected by JC-10, mitochondrial membrane potential was quantified by red-green fluorescence ratio. ∗∗∗ p < 0.001 (n = 8) ( G ); Representative TEM images of RAECs mitochondria, the arrows indicate mitochondria ( H ); ( I–O ) RAECs were transfected with MPC1 siRNA to knock down MPC1. Representative images of immunofluorescence analysis of eNOS in RAECs after knocking down MPC1 ( I ); The protein expression of eNOS, p-eNOS, ZO-1, Occludin and VE-Cadherin was detected. ∗∗∗ p < 0.001 (n = 4) ( J ); The protein and mRNA levels of MCT4 and MPC1 were detected. ∗∗ p < 0.01, ∗∗∗ p < 0.001 (n = 4) ( K ); Lactate content in the supernatant of RAECs culture. ∗∗ p < 0.01 (n = 3) ( L ); Lactate content in the cytosol of RAECs, normalized using intracellular protein content. ∗ p < 0.05 (n = 3) ( M ); The oxygen consumption rate (OCR) of RAECs after MPC1 knockdown was analyzed by Seahorse XF analyzer, and the ATP production was normalized using intracellular protein content. ∗∗ p < 0.01 (n = 3) ( N ). Representative images of the mitochondrial membrane potential of RAECs detected by JC-10 after knocking down MPC1, and mitochondrial membrane potential was quantified by red-green fluorescence ratio. ∗∗∗ p < 0.001 (n = 7) ( O ).

Article Snippet: Mouse MCT4 (sense: CACGGCAGGUUUCAUAACATT, antisense: UGUUA UGAAACCUGCCGUGTT), MPC1 (sense: GCUAUUCUCUGACAUUCAUTT, antisense: AUGAAUGUCAGAGAAUAGCTT), PKM2 (sense: GCCACAGAAA GCUUUGCAUTT, antisense: AUGCAAAGCUUUCUGUGGCTT) and control small interfering RNA (siRNA) were obtained from GenePharma (Shanghai, China).

Techniques: Blocking Assay, Immunofluorescence, Expressing, Membrane, Fluorescence, Transfection, Knockdown

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: 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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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: For the si MPC1 cell line, H1299 cells (1 × 10 6 /well) were infected with siRNAs targeting MPC1 (GenePharma, Shanghai, China).

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