Journal: The Journal of international medical research

Article Title: MYL9 promotes squamous cervical cancer migration and invasion by enhancing aerobic glycolysis.

doi: 10.1177/03000605231208582

Figure Lengend Snippet: Figure 3. MYL9 promoted the progression of SCC by enhancing aerobic glycolysis. (a) MYL9 knockdown decreased the Lactate production of SCC. (b) MYL9 knockdown inhibited the expression of GLUT1, HK2, and LDHA and (c) MYL9 knockdown inactivated the JAK2/STAT3 pathway. MYL9, myosin light chain 9; SCC, squamous cervical cancer; GLUT1, glucose transporter 1; HK2, hexokinase II; LDHA, lactate dehydrogenase A.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in TBS containing Tween-20 (TBST) for 1 hour at 37 C and incubated with primary antibodies againstMYL9 (mouse, 60233-1-Ig, Proteintech, Rosemont, IL, USA), GLUT1 (rabbit, 21829-1-AP, Proteintech), HK2 (mouse, 66974-1-Ig, Proteintech), LDHA (rabbit, 19987-1-AP, Proteintech), b-tubulin (mouse, Proteintech), and GAPDH (mouse, CW0100M, CWBIO, Beijing, China) overnight at 4 C. Following three washes with TBST, the membrane was incubated with horseradish peroxidase-conjugated goat antirabbit or anti-mouse IgG antibody at 37 C for 1 hour.

Techniques: Knockdown, Expressing

Journal: Journal of translational medicine

Article Title: TIMELESS promotes reprogramming of glucose metabolism in oral squamous cell carcinoma.

doi: 10.1186/s12967-023-04791-3

Figure Lengend Snippet: Fig. 5 TIMELESS augments glycolysis via upregulation of key metabolic Molecules. A Correlation analysis between TIMELESS expression and SIRT1, HIF1A and MYC in HNSC tissues form TCGA database. B Correlation analysis between TIMELESS mRNA levels and SIRT1 mRNA levels in 33 OSCC tissues. C qRT-PCR analysis of the expression levels of TIMELESS and key glycolysis genes in OSCC cells. D Western blot analysis of the expression levels of TIMELESS and key glycolysis genes in OSCC cells. E IHC staining of TIMELESS, SIRT1, HK2, PKM2, LDHA and GLUT1 in nude mice tumors. F The transfection efficiency was examined with western blot. G The level of glucose uptake was examined. H Lactate production was examined. I Cell medium pH. J Oxygen consumption level of cell. Data shown were the mean ± S.E.M. from three independent experiments. *P < 0.05; **P < 0.01

Article Snippet: For the immunohistochemistry assays, we used the Ki67 antibody (AF0198) from Affinity Biosciences, and the PCNA antibody (60097-1-Ig), SIRT1 antibody (13161- 1-AP), HK2 antibody (66974-1-Ig), PKM2 antibody (60268-1-Ig), GLUT1 antibody (66290-1-Ig), LDHA antibody (66287-1-Ig), and TIMELESS antibody (14421-1- AP) from Proteintech Group, Inc.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Transfection

Journal: Investigative Ophthalmology & Visual Science

Article Title: IL-6-Derived Autocrine Lactate Promotes Immune Escape of Uveal Melanoma

doi: 10.1167/iovs.65.3.37

Figure Lengend Snippet: IL-6 increased glycolysis and lactate production in C918 cells. (A) Distribution of differentially expressed transcripts between IL-6/C918 and C918 cells identified in the glycolysis/gluconeogenesis pathway (KEGG pathway), based on transcriptome data; red = upregulated and blue = downregulated. Pathway with blue shading indicated the glycolysis process, and pathway with red shading indicated the pyruvate oxidation process. (B–D) Relative mRNA levels of PFKP (B), PKM (C), and LDHA (D) in IL-6/C918 and C918 cells determined by qPCR. (E) PFKP, PKM, and LDHA protein levels in IL-6/C918 and C918 cells detected by Western blotting. (F) Analysis of TCGA uveal melanoma database using GEPIA2 ( http://gepia2.cancer-pku.cn/#correlation ) showing correlations between IL-6 and PKM or LDHA mRNA levels. (G) Extracellular acidification rate (ECAR) of IL-6/C918 and C918 cells. (H) Lactate concentrations in the culture medium of IL-6/C918 and C918 cells. The results of three independent experiments are presented as mean ± standard deviation.

Article Snippet: Antibodies against PFKP (sc-514824), PKM (sc-365684), LDHA (sc-137243), PD-L1 (sc-293425), HLA-E (sc-51621), and ubiquitin (sc-8017) were purchased from Santa Cruz Biotechnology (SantaCruz, CA, USA).

Techniques: Western Blot, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: IL-6-Derived Autocrine Lactate Promotes Immune Escape of Uveal Melanoma

doi: 10.1167/iovs.65.3.37

Figure Lengend Snippet: IL-6-induced lactate increased PD-L1 and HLA-E protein levels and immune escape in C918 cells. (A and B) PD-L1 and HLA-E protein and relative mRNA levels in IL-6/C918 cells with PKM or LDHA knocked down examined by Western blotting and qPCR. (C and D) PD-L1 and HLA-E protein and relative mRNA levels in IL-6/C918 cells treated with 50 nM sodium oxamate (SO) examined by Western blotting and qPCR. (E and F) PD-L1 and HLA-E protein and relative mRNA levels examined by Western blotting and qPCR in C918 cells treated with 20 mM lactate. (G) Apoptosis level of IL-6/C918 cells pretreated with 50 nM SO after co-culture with TALL 104 and NK-92MI cells. (H) Apoptosis level of C918 cells pretreated with 20 mM lactate after co-culture with TALL-104 and NK-92MI cells. The results of three independent experiments are presented as mean ± standard deviation.

Article Snippet: Antibodies against PFKP (sc-514824), PKM (sc-365684), LDHA (sc-137243), PD-L1 (sc-293425), HLA-E (sc-51621), and ubiquitin (sc-8017) were purchased from Santa Cruz Biotechnology (SantaCruz, CA, USA).

Techniques: Western Blot, Co-Culture Assay, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: IL-6-Derived Autocrine Lactate Promotes Immune Escape of Uveal Melanoma

doi: 10.1167/iovs.65.3.37

Figure Lengend Snippet: LDHA protein levels were positively correlated with those of PD-L1 in uveal melanoma tissues. (A) Two representative cases of low and high expression of PKM, LDHA, PD-L1, and HLA-E in clinical UM tissues. (B) The relationships of the integrated fluorescence density of PKM, LDHA, PD-L1, and HLA-E.

Article Snippet: Antibodies against PFKP (sc-514824), PKM (sc-365684), LDHA (sc-137243), PD-L1 (sc-293425), HLA-E (sc-51621), and ubiquitin (sc-8017) were purchased from Santa Cruz Biotechnology (SantaCruz, CA, USA).

Techniques: Expressing, Fluorescence

Journal: Cell communication and signaling : CCS

Article Title: Blockade of glycolysis-dependent contraction by oroxylin a via inhibition of lactate dehydrogenase-a in hepatic stellate cells.

doi: 10.1186/s12964-019-0324-8

Figure Lengend Snippet: Fig. 5 Oroxylin A inhibition of LDH-A led to blockade of aerobic glycolysis in HSCs. LX2 cells were treated with oroxylin A and/or galloflavin at indicated concentrations, or transfected with LDH-A siRNA for 24 h. a Measurements of glucose uptake. b Measurements of glucose consumption indicated by GOD activity. c Measurements of intracellular lactate levels. d Real-time PCR analyses of mRNA expression of HK2, PFK1 and PKM2. e Western blot analyses of protein expression of HK2, PFK1 and PKM2 with quantification. f Measurements of intracellular HK2 enzyme activity. g Measurements of intracellular PFK1 enzyme activity. h Measurements of intracellular PKM2 enzyme activity. i Real-time PCR analyses of mRNA expression of LDH-A. j Western blot analyses of protein expression of LDH-A with quantification. k Real-time PCR analyses of mRNA expression of HK2, PFK1 and PKM2. For statistical significance of panels a-h: *p < 0.05 vs. control, **p < 0.01 vs. control, ***p < 0.001 vs. control. For statistical significance of panels i-k: *p < 0.05 vs. control siRNA, **p < 0.01 vs. control siRNA, ***p < 0.001 vs. control siRNA

Article Snippet: LDH-A siRNA (sc-43,893) and control siRNA (sc-37,007) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Inhibition, Transfection, Activity Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: Cell communication and signaling : CCS

Article Title: Blockade of glycolysis-dependent contraction by oroxylin a via inhibition of lactate dehydrogenase-a in hepatic stellate cells.

doi: 10.1186/s12964-019-0324-8

Figure Lengend Snippet: Fig. 6 Oroxylin A inhibition of LDH-A led to reduction of HSC contraction. LX2 cells were treated with oroxylin A and/or galloflavin at indicated concentrations, or transfected with LDH-A siRNA for 24 h. a Collagen gel contraction assays with quantification. b Cytoskeleton fluorescence staining, scale bar: 20 μm. c Western blot analyses of MLC2 phosphorylation with quantification. d Immunofluorescence analyses of MLC2 phosphorylation, scale bar: 20 μm. e Cytoskeleton fluorescence staining, scale bar: 20 μm. f Western blot analyses of MLC2 phosphorylation with quantification. For statistical significance of panels a-d: *p < 0.05 vs. control, **p < 0.01 vs. control. For statistical significance of panels e-f: **p < 0.01 vs. control siRNA

Article Snippet: LDH-A siRNA (sc-43,893) and control siRNA (sc-37,007) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Inhibition, Transfection, Fluorescence, Staining, Western Blot, Phospho-proteomics, Immunofluorescence, Control

Journal: Traffic (Copenhagen, Denmark)

Article Title: Assembly of Peroxisomal Membrane Proteins via the Direct Pex19p-Pex3p Pathway.

doi: 10.1111/tra.12376

Figure Lengend Snippet: Figure 2: Pex19p forms soluble complexes with PMP22 in the cytosol. A) Overexpression of Pex19p stabilizes the coexpressed PMP22 in the cytosol. CHO-K1 cells expressing PMP22-HA alone (lanes 1–3) or together with FLAG-Pex19p (lanes 4–6) were resuspended with a buffer containing 100 μg/mL of digitonin to permeabilize the plasma membrane, and analyzed directly (T; lanes 1 and 4) or after separation into cytosolic (C; lanes 2 and 5) and organelle-containing (O; lanes 3 and 6) fractions. Equal aliquots of each fraction were analyzed by SDS–PAGE and immunoblotting with the indicated antibodies. Lactate dehydrogenase (LDH) and Pex14p are markers for cytosolic and organelle fractions, respectively. B) Pex19p binds to PMP22 in the cytosol. Cytosolic fraction from CHO-K1 cells overexpressing FLAG-Pex19p or PMP22-HA plus FLAG-Pex19p was subjected to immunoprecipitation (IP) with pre-immune serum (lanes 3 and 4) or anti-HA antibody (lanes 5 and 6). Input (10% of that used for IP; lanes 1 and 2) and immunoprecipitates were analyzed by SDS–PAGE and immunoblotting using the indicated antibodies. C) Farnesylation of Pex19p increases the binding affinity of Pex19p toward PMP22. Whole-cell lysates (WCLs) from CHO-K1 cells each expressing PMP22-HA together with epitope-tagged Pex19p variants, including Myc-Pex19p (WT, lanes 1 and 5), Myc-Pex19p-C296S (C296S, lanes 2 and 6), FLAG-Pex19p (WT, lanes 3 and 7) and FLAG-Pex19p-C296S (C296S, lanes 4 and 8) were subjected to IP with anti-FLAG agarose beads. Input (10% of that used for IP; lanes 1–4) and immunoprecipitates (lanes 5–8) were analyzed by SDS–PAGE and immunoblotting with anti-HA, anti-Pex19p and anti-α-tubulin antibodies. Solid and open arrowheads show unmodified and farnesylated epitope-tagged Pex19p, respectively. Tubulin was analyzed as a loading control.

Article Snippet: Rabbit antibodies against FLAG (Sigma-Aldrich) and GFP (Medical and Biological Laboratories); mouse monoclonal antibodies against FLAG (M2; Sigma-Aldrich), HA (16B12; Covance), c-Myc (9E10; Santa Cruz Biotechnology, Inc.), Pex19p (BD), α-tubulin (DM1A; Abcam), Tom20 (F-10; Santa Cruz Biotechnology, Inc.) and GFP (B-2; Santa Cruz Biotechnology, Inc.); and goat anti-lactate dehydrogenase (LDH) antibody (Rockland Immunochemicals) were purchased.

Techniques: Over Expression, Expressing, Clinical Proteomics, Membrane, SDS Page, Western Blot, Immunoprecipitation, Binding Assay, Control

Journal: Traffic (Copenhagen, Denmark)

Article Title: Assembly of Peroxisomal Membrane Proteins via the Direct Pex19p-Pex3p Pathway.

doi: 10.1111/tra.12376

Figure Lengend Snippet: Figure 4: Pex3p-Pex19p interaction is essential for peroxisomal targeting of various PMPs. A–C) Pex19p-PMP complexes are targeted to peroxisomes in a manner dependent on Pex3p-Pex19p interaction. Left, cytosolic fractions from CHO-K1 cells expressing HA-Pex16p (A), PMP34-HA (B) or PMP70-HA (C), together with either FLAG-Pex19p or FLAG-Pex19pΔN39 were subjected to the in vitro import assay as described in Figure 3A. After the import reaction, cells were stained with anti-HA and anti-PTS1 antibodies. Merged views are shown on the right (c and f). Scale bars, 10 μm. Insets show threefold enlarged images of the boxed areas. Right, cells that were incubated with cytosolic fractions containing the indicated Pex19p-PMP complexes (A–C, a–c) were treated with 0.1 M Na2CO3 and ultracentrifuged. Equal aliquots of soluble (S, lane 1) and membrane (P, lane 2) fractions were analyzed by SDS–PAGE and immunoblotting with anti-HA, anti-Pex14p and anti-AOx antibodies. For AOx, only the AOx-B chain was shown. D) Cytosolic fractions used in (A–C) were analyzed by SDS–PAGE and immunoblotting with anti-HA, anti-Pex19p and anti-lactate dehydrogenase (LDH) antibodies. Full, FLAG-Pex19p; ΔN39, FLAG-Pex19pΔN39. Solid and open arrowheads indicate unmodified and farnesylated FLAG-Pex19p (full and ΔN39), respectively.

Article Snippet: Rabbit antibodies against FLAG (Sigma-Aldrich) and GFP (Medical and Biological Laboratories); mouse monoclonal antibodies against FLAG (M2; Sigma-Aldrich), HA (16B12; Covance), c-Myc (9E10; Santa Cruz Biotechnology, Inc.), Pex19p (BD), α-tubulin (DM1A; Abcam), Tom20 (F-10; Santa Cruz Biotechnology, Inc.) and GFP (B-2; Santa Cruz Biotechnology, Inc.); and goat anti-lactate dehydrogenase (LDH) antibody (Rockland Immunochemicals) were purchased.

Techniques: Expressing, In Vitro, Staining, Incubation, Membrane, SDS Page, Western Blot

Journal: The FEBS journal

Article Title: Knockdown of lactate dehydrogenase A suppresses tumor growth and metastasis of human hepatocellular carcinoma.

doi: 10.1111/j.1742-4658.2012.08748.x

Figure Lengend Snippet: Fig. 1. Elevated expression of LDHA correlates with increased metastatic potential in HCC cells. (A) Protein expression of LDHA in one normal liver cell line (L–02) and five cancer cell lines (HCCLM3, MHCC97–L, MHCC97–H, HepG2 and Hep3B) was examined by western blot. (B) Inhibitory effects of lentiviral-mediated RNAi on LDHA expression. Relative LDHA mRNA levels were analysed by quantitative RT–PCR in cell lines transiently transfected with or stably expressing LDHA shRNA. (C) Western blot analysis was performed on whole-cell lysates prepared from cell lines transiently transfected with or stably expressing LDHA shRNA. (D) LDHA activity in knockdown clones, expressed as percentage of enzyme activity in parental HCCLM3 cells. b–actin was used as a normalization control. The data shown are representative of three separate experiments.

Article Snippet: shRNA lentiviral particles targeting LDHA (LDHA shRNA Lenti sc–43893–v), and the appropriate control shRNA lentiviral particles (control shRNA Lenti sc–108080), were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and transfected into the HCCLM3 cell line according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Transfection, Stable Transfection, shRNA, Activity Assay, Knockdown, Clone Assay, Control

Journal: The FEBS journal

Article Title: Knockdown of lactate dehydrogenase A suppresses tumor growth and metastasis of human hepatocellular carcinoma.

doi: 10.1111/j.1742-4658.2012.08748.x

Figure Lengend Snippet: Fig. 2. Functional role of LDHA in in vitro proliferation of HCCLM3. (A) Cell viability was determined by cell counting after treatment of HCCLM3 cells with LDHA shRNA Lenti. (B) Comparison of glucose consumption rate across LDHA knockdown cells and control cells. (C) Lactate production rate detected under the same experimental conditions as in (A). (D) Oxygen consumption of HCCLM3 cells was determined by the use of a Clark-type oxygen electrode at 72 h post-transfection with shRNAs. (E) Quantification of ATP levels in cells grown under normoxic conditions (white bars) and hypoxic conditions (black bars) for 48 h using a luciferin/luciferase-based assay. (F) ROS measurement by H2DCFDA in control cells and LDHA-deficient cell lines in the presence and absence of N–acetylcysteine. (G) The intracellular free Ca2+ concentration ([Ca2+]i) was determined using FLUO–3AM under confocal microscope. (H) Assessment of mitochondrial membrane potential using JC–1 fluorescent dye by confocal microscopy. (I) Cytochrome c release from the mitochondria in LDHA shRNA Lenti cells. Cells were treated with LDHA shRNA Lenti for 48 h, and 5 lg mitochondria-free cytosolic protein fraction was analyzed for cytochrome c by ELISA. The results represent the mean ± SEM of three independent experiments. *P < 0.05 versus HCCLM3 and control shRNA Lenti. **P < 0.05 versus cells without N–acetylcysteine (NAC).

Article Snippet: shRNA lentiviral particles targeting LDHA (LDHA shRNA Lenti sc–43893–v), and the appropriate control shRNA lentiviral particles (control shRNA Lenti sc–108080), were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and transfected into the HCCLM3 cell line according to the manufacturer’s instructions.

Techniques: Functional Assay, In Vitro, Cell Counting, shRNA, Comparison, Knockdown, Control, Transfection, Luciferase, Concentration Assay, Microscopy, Membrane, Confocal Microscopy, Enzyme-linked Immunosorbent Assay

Journal: The FEBS journal

Article Title: Knockdown of lactate dehydrogenase A suppresses tumor growth and metastasis of human hepatocellular carcinoma.

doi: 10.1111/j.1742-4658.2012.08748.x

Figure Lengend Snippet: Fig. 3. Induction of apoptosis in HCCLM3 by LDHA RNAi. (A) After 72 h transfection, cells were isolated and immunostained with annexin V-FITC/propidium iodide (PI) and analysed by fluorescence-activated cell sorting (FACS). (B) Time-dependent LDHA shRNA Lenti-induced apoptosis in HCCLM3 was determined using caspase–3 by fluorescence-activated cell sorting. (C) LDHA knockdown-induced apoptosis of HCCLM3 using caspase–3 staining was studied after exposure to 1 mM of the antioxidant N–acetylcysteine for 18 h. (D) Proposed model for LDHA inhibition-induced apoptosis in human HCC cells. LDHA inhibition induced production of ROS and cytosolic Ca2+, and decreased the inner mitochondrial membrane potential (DΨm), leading to caspase–9 and caspase–3 activation before causing apoptosis in HCCLM3 cells. *P < 0.05 versus HCCLM3 and control shRNA Lenti.

Article Snippet: shRNA lentiviral particles targeting LDHA (LDHA shRNA Lenti sc–43893–v), and the appropriate control shRNA lentiviral particles (control shRNA Lenti sc–108080), were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and transfected into the HCCLM3 cell line according to the manufacturer’s instructions.

Techniques: Transfection, Isolation, FACS, shRNA, Knockdown, Staining, Inhibition, Membrane, Activation Assay, Control

Journal: The Journal of Biological Chemistry

Article Title: Leptin enhances hypothalamic lactate dehydrogenase A (LDHA)–dependent glucose sensing to lower glucose production in high-fat–fed rats

doi: 10.1074/jbc.RA117.000838

Figure Lengend Snippet: High-fat feeding impairs hypothalamic glucose sensing. A, experimental protocol. B, cumulative food intake. *, p < 0.05 compared with regular chow (RC)–fed. Body weight (C), fat and lean mass (D), and free water and total water mass (E) of HFD (n = 7)- versus RC (n = 5)-fed rats are shown. F, glucose infusion rate. G, glucose production in basal (white bars) and clamp (black bars) conditions. H, glucose uptake during the clamps. n = 7 for RC-MBH saline, n = 6 for RC-MBH glucose, n = 7 for HFD-MBH saline, and n = 7 for HFD-MBH glucose. Error bars represent S.E. *, p < 0.05 compared with RC-saline, HFD-saline, and HFD-glucose. LV, lentivirus.

Article Snippet: Immediately following brain surgery, a group of rats received 3 μl of lentivirus expressing shRNA to LDHA (1.0 × 10 6 infectious units; sc-270631-V, Santa Cruz Biotechnology, Inc., Dallas, TX) or a mismatch sequence as a control (1.0 × 10 6 infectious units; sc-108080, Santa Cruz Biotechnology, Inc.) through each side of the MBH catheters that target the arcuate nucleus ( 29 , 30 ).

Techniques: Saline

Journal: The Journal of Biological Chemistry

Article Title: Leptin enhances hypothalamic lactate dehydrogenase A (LDHA)–dependent glucose sensing to lower glucose production in high-fat–fed rats

doi: 10.1074/jbc.RA117.000838

Figure Lengend Snippet: Molecular knockdown of hypothalamic LDH negates leptin's impact on glucose sensing. A, representative Western blot of LDHA and LDHB in the MBH of rats that received MBH injections of either MM (n = 4) or LDHA shRNA lentivirus (n = 5). Quantification of LDHA or LDHB levels normalized to β-tubulin expressed as -fold change over MM is shown below. Error bars represent S.E. *, p < 0.05 compared with MM. Glucose infusion rate (B), glucose production in basal (white bars) and clamp (black bars) conditions (C), and glucose uptake during the clamps (D) (n = 7 per group) are shown. Error bars represent S.E. *, p < 0.05 compared with RC-MBH saline and RC-MBH glucose + LDHA shRNA; †, p < 0.05 compared with HFD-MBH saline + MM, HFD-MBH glucose + MM, HFD-MBH glucose + LDHA shRNA, and HFD-MBH glucose + leptin + LDHA shRNA.

Article Snippet: Immediately following brain surgery, a group of rats received 3 μl of lentivirus expressing shRNA to LDHA (1.0 × 10 6 infectious units; sc-270631-V, Santa Cruz Biotechnology, Inc., Dallas, TX) or a mismatch sequence as a control (1.0 × 10 6 infectious units; sc-108080, Santa Cruz Biotechnology, Inc.) through each side of the MBH catheters that target the arcuate nucleus ( 29 , 30 ).

Techniques: Knockdown, Western Blot, shRNA, Saline

Journal: The Journal of Biological Chemistry

Article Title: Leptin enhances hypothalamic lactate dehydrogenase A (LDHA)–dependent glucose sensing to lower glucose production in high-fat–fed rats

doi: 10.1074/jbc.RA117.000838

Figure Lengend Snippet: Plasma insulin and glucose concentrations during the basal and clamp periods relating to Errors represent S.E. (basal, 60–90 min; clamp, 180–210 min).

Article Snippet: Immediately following brain surgery, a group of rats received 3 μl of lentivirus expressing shRNA to LDHA (1.0 × 10 6 infectious units; sc-270631-V, Santa Cruz Biotechnology, Inc., Dallas, TX) or a mismatch sequence as a control (1.0 × 10 6 infectious units; sc-108080, Santa Cruz Biotechnology, Inc.) through each side of the MBH catheters that target the arcuate nucleus ( 29 , 30 ).

Techniques: Clinical Proteomics, shRNA, Saline