Journal: Science Advances

Article Title: Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission

doi: 10.1126/sciadv.adp7423

Figure Lengend Snippet: ( A and C ) Co-immunostaining of hippocampal neurons with antibodies against MPC1 (A) or MPC2 (C) and the presynaptic protein vGLUT1. Scale bars, 10 μm. ( B and D ) Magnification of the boxed areas in (A) and (C). Colocalization denoted with arrowheads. Scale bars, 10 μm (B) and 5 μm (D). ( E ) Representative images of axonal mitochondria expressing a mitochondrial pyruvate sensor, CamKII-mtPyronicSF, showing an increase in fluorescence intensity with perfusion of 10 mM extracellular pyruvate compared to 0 mM. Scale bar, 10 μm. ( F ) Average traces of CamKII-mtPyronicSF (Δ F / F 0 ) showing inhibition of mitochondrial pyruvate uptake in neurons expressing shRNA against mpc1 (MPC1 KD). F: fluorescence intensity. n = 12 to 18 (neurons). ( G ) Maximal CamKII-mtPyronicSF fluorescence response in 10 mM pyruvate as determined from traces in (F). ( H ) Schematic of the pyruvate sensor CaMKII-mtPyronicSF and the ATP indicator Syn-ATP expressed in nerve terminals. ( I ) Presynaptic ATP traces in terminals supplied with lactate and pyruvate (lac/pyr) after incubation with the MPC inhibitor UK5099 (100 μM), normalized to preincubation levels. n = 9 (neurons). ( J ) Presynaptic ATP level after application of MPC inhibitor UK5099 normalized to ATP level before. The box-whisker plots denote median (line), 25th to 75th percentile (box), and minimum-maximum (whiskers). Man-Whitney U test (G) and one sample t test (J). Error bars are SEM.

Article Snippet: Human Mpc1 TaqMan Probe , Thermo Fisher Scientific , Catalog no. Hs00211484_m1.

Techniques: Immunostaining, Expressing, Fluorescence, Inhibition, shRNA, Incubation, Whisker Assay

Journal: Science Advances

Article Title: Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission

doi: 10.1126/sciadv.adp7423

Figure Lengend Snippet: ( A ) Representative images of an HEK293 cell expressing mtPyronicSF in media containing 0 and 10 mM pyruvate. ( B ) Average traces of mtPyronicSF showing differential mitochondrial pyruvate accumulation in control HEK293 cells and cells expressing shRNA against sirt3 (Sirt3 KD) or mpc1 (MPC1 KD). ( C ) Peak values of mtPyronicSF (Δ F / F 0 ) in response to 10 mM pyruvate, as determined from traces in (B). n = 24 to 57 fields of view (FOV), five to six cells per FOV. ( D ) Immunoprecipitation (IP) of endogenous MPC1 from Sirt3 +/+ and Sirt3 −/− mouse brain lysates with antibodies against MPC1 and rabbit immunoglobulin G (IgG) (control) followed by immunoblotted (IB) with antibodies against MPC1 (top) and Ac-K (bottom). Asterix denotes a nonspecific band in the IgG immunoprecipitate. ( E ) Intensity of Ac-K band normalized to MPC1 band from (D), expressed relative to Sirt3 +/+ control. n = 4 (IP blots). Raw intensities of Ac-K normalized to MPC1 in Sirt3 +/+ and Sirt3 −/− brains were used for statistical comparison. ( F ) Alignment of rat ( Rattus norvegicus, R. n. ), mouse ( Mus musculus, M. m. ), and human ( Homo sapiens, H. s. ) MPC1 protein sequences showing conservation of an acetylation motif (boxed in blue). A nonacetylatable form was constructed by mutation of K45/46 to A45/A46. ( G ) Immunoprecipitation of wild-type (WT) MPC1 or acetyl mutant (MPC1-AA) from Sirt3-deficient (KD) HEK293 cells, immunoblotted for FLAG (top) and Ac-K (bottom). ( H ) Intensity of Ac-K band normalized to FLAG intensity from panel G, expressed relative to WT-MPC1. n = 6 (IP blots). Kruskal-Wallis test (C), paired t test (E), and one sample t test (H). Error bars are SEM. IB, immunoblot. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Human Mpc1 TaqMan Probe , Thermo Fisher Scientific , Catalog no. Hs00211484_m1.

Techniques: Expressing, Control, shRNA, Immunoprecipitation, Comparison, Construct, Mutagenesis, Western Blot

Journal: Science Advances

Article Title: Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission

doi: 10.1126/sciadv.adp7423

Figure Lengend Snippet: ( A ) Schematic of a HEK293 cell expressing mtPyronicSF. ( B ) Average traces of mtPyronicSF expressed in HEK293 cells showing differential mitochondrial pyruvate accumulation in control, MPC1 KD, MPC1 KD + WT MPC1, or MPC1 KD + MPC1-QQ (acetyl mimetic mutant). Control and MPC1 KD traces are the same as in . ( C ) Peak values of mtPyronicSF (Δ F / F 0 ) in response to perfusion of 10 mM pyruvate, as determined from traces in (B). n = 22 to 57 FOV, five to six cells per FOV. ( D ) Schematic of a hippocampal neuron-expressing vGLUT1-pH. ( E ) Sample normalized vGLUT1-pH traces in hippocampal terminals electrically stimulated with 100 AP at 10 Hz in control neurons, or neurons expressing MPC1 KD, MPC1 KD + WT MPC1, or MPC1 KD + MPC1-QQ (acetyl mimetic mutant). ( F ) SV retrieval quantified as fractional retrieval block calculated from traces in (E). n = 16 to 81 (neurons). Black bar denotes electrical stimulation. Kruskal-Wallis test [(C) and (F)]. Error bars are SEM. *** P < 0.001; **** P < 0.0001.

Article Snippet: Human Mpc1 TaqMan Probe , Thermo Fisher Scientific , Catalog no. Hs00211484_m1.

Techniques: Expressing, Control, Mutagenesis, Blocking Assay

Journal: Science Advances

Article Title: Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission

doi: 10.1126/sciadv.adp7423

Figure Lengend Snippet: Key resource table. ATCC, American Type Culture Collection; IgG, immunoglobulin G; N/A, not applicable.

Article Snippet: Human Mpc1 TaqMan Probe , Thermo Fisher Scientific , Catalog no. Hs00211484_m1.

Techniques: Western Blot, shRNA, Inhibition, Magnetic Beads, Plasmid Preparation, Sequencing, CRISPR, Software

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: 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, Injection

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: Sequences of the sense strands of siRNAs targeting different genes.

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:

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: bioRxiv

Article Title: Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

doi: 10.1101/2023.01.31.526492

Figure Lengend Snippet: (A) Bar graph comparing kidney Mpc1 mRNA levels after vehicle treatment or cisplatin-, ischemia reperfusion (IR)-, and rhabdomyolysis (Rhabdo)-induced AKIs. Samples were collected 72 hours after cisplatin injury and 24 hours after IR and rhabdomyolysis injuries. (n = 6/group; *** p < 0.001 by unpaired t test with Welch’s correction). (B) Representative Western blot of kidney MPC1 protein abundance after AKI. Samples were collected 72 hours after cisplatin injury and 24 hours after IR and rhabdomyolysis injuries. β-ACTIN was blotted as a loading control. (C) Representative immunostaining images of MPC1 (red), lotus tetragonolobus lectin (LTL, green, proximal tubule marker), or peanut agglutinin (PNA, green, distal tubule marker), and DAPI (blue) in whole kidney, outer cortex (OC) and cortico-medullary junction (CM) kidney sections 30 hours following vehicle treatment or rhabdomyolysis-induced AKI. (Images captured at 15x magnification; whole kidney scale bar = 1,000 μm; OC and CM scale bar = 50 μm). ( D ) Representative fluorescence image of kidney sections of mT/mG/Ggt1-Cre mice confirming GFP+ renal tubular epithelial cells (green, #, GFP) and tdTomato+ non-RTEC cells (red, *, tdT) stained with Dapi (blue). (Scale bar = 100 μm). (E) Bar graph comparing Mpc1 mRNA levels in flow-sorted Non-RTEC (tdTomato+) and RTEC (GFP+) cells 24 hour after vehicle treatment or rhadbomyolysis-induced AKI. (n = 5/group, ***p < 0.001 by unpaired t test with Welch’s correction). (F) Representative Western blot of MPC1 and VDAC protein abundance in flow-sorted Non-RTEC (tdTomato+) and RTEC (GFP+) cells 24 hour after AKI. β-ACTIN was blotted as a loading control. Data are presented as means ± SEM.

Article Snippet: The MPC1 f/f mice were bred with mice that express Cre recombinase under the control of the tubule specific Pax8 Cre promoter (Jackson Laboratory, 028196) ( , ).

Techniques: Western Blot, Control, Immunostaining, Marker, Fluorescence, Staining

Journal: bioRxiv

Article Title: Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

doi: 10.1101/2023.01.31.526492

Figure Lengend Snippet: (A) Schematic illustrating the generation of tubular Mpc1 null allele, MPC TubKO mice (TubKO). (B-C) Bar graphs showing body weights ( B ) and serum cystatin C concentration ( C ) in WT and MPC TubKO mice. (n = 5/group, 8-week-old mice). (D) Bar graph comparing mouse kidney Mpc1 mRNA levels in WT and MPC TubKO mice. (n = 4/group, 7 – 12-week-old mice, ** p < 0.01 by unpaired t test with Welch’s correction). (E-G) Representative Western blot of kidney MPC1 and MPC2 protein abundance ( E ) and quantification of normalized MPC1 ( F ) and MPC2 ( G ) levels in WT and MPC TubKO mice. Tubulin was blotted as loading control and used as the protein quantification normalizer. (n = 4 – 6/group, 7 – 12-week-old mice, *** p < 0.001 and ** p < 0.01 by by unpaired t test with Welch’s correction). (H) Representative immunostaining images of kidney MPC1 (green) and lotus tetragonolobus lectin (LTL, green, proximal tubule marker) or peanut agglutinin (PNA, green, distal tubule marker) in whole kidney (WK), outer-cortex (OC), and cortico-medullary junction (CM) in WT and MPC TubKO mice. (Images taken at 4x (WK) or 20x (OC and CM) magnification, scale bar = 500 μm). Data are presented as means ± SEM.

Article Snippet: The MPC1 f/f mice were bred with mice that express Cre recombinase under the control of the tubule specific Pax8 Cre promoter (Jackson Laboratory, 028196) ( , ).

Techniques: Concentration Assay, Western Blot, Control, Immunostaining, Marker

Journal: bioRxiv

Article Title: Tubular Mitochondrial Pyruvate Carrier Disruption Elicits Redox Adaptations that Protect from Acute Kidney Injury

doi: 10.1101/2023.01.31.526492

Figure Lengend Snippet: ( A ) Line graph showing the survival curve of WT and MPC TubKO mice following rhabdomyolysis (Rhabdo)-induced AKI. (n = 10 – 11/group, 8 – 12-week-old mice, * p < 0.05 by Mantel-Cox log-rank test). ( B-C ) Bar graphs showing serum cystatin C ( C ), and blood urea nitrogen ( D , BUN) levels prior to (D0) and on day 1 (D1, 24-hours) and day 2 (D2, 48 hours) after vehicle treatment or rhabdomyolysis-induced AKI in WT and MPC TubKO mice. (n = 10 – 11/group, 8 – 12-week-old mice, * p< 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA followed by Turkey’s multiple comparison tests). ( E-F ) Bar graphs showing kidney Ngal ( D ) and Kim1 ( E ) mRNA levels one day (24 hours) after vehicle treatment or rhabdomyolysis-induced AKI in WT and MPC TubKO mice. (n = 4/group for vehicle treatment, n = 12 – 13/group for Rhabdo, 8 – 12-week-old mice, * p < 0.05 by two-way ANOVA with Tukey’s multiple comparison test). ( G-H ) Bar graphs showing quantification of histologically assessed tubular injury score ( F ) and tunel positive tubular cells ( G ) one day (24 hours) after vehicle treatment or rhabdomyolysis-induced AKI in WT and MPC TubKO mice. (n = 4/group for vehicle treatment, n = 12 – 13/group for Rhabdo, 8 – 12-week-old mice, ** p < 0.01 and *** p < 0.001 by two-way ANOVA with Tukey’s multiple comparison test). ( H-J ) Heatmaps showing Spearman correlation between variables analyzed following vehicle treatment or rhabdomyolysis-induced AKI. Correlation calculated in WT mice comparing Mpc1 mRNA levels, tubular injury, Ngal and Kim1 mRNA levels, tunel score, and tubular GSH with AKI ( H ). Spearman correlation performed in WT ( I ) and MPC TubKO ( J ) mice comparing GSH and antioxidant defense system markers following rhabdomyolysis-induced AKI. Data are presented as means ± SEM.

Article Snippet: The MPC1 f/f mice were bred with mice that express Cre recombinase under the control of the tubule specific Pax8 Cre promoter (Jackson Laboratory, 028196) ( , ).

Techniques: Comparison, TUNEL Assay