Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Primer sequences for real-time PCR amplification.

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Real-time Polymerase Chain Reaction, Amplification

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the mRNA expression levels of Zn, amino acid and small peptide transporters in the jejunum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the mRNA expression levels of Zn, amino acid and small peptide transporters in the ileum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the protein expression levels of Zn, amino acid and small peptide transporters in the jejunum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Representative immunoblots of ZnT1, ZnT4, ZnT5, ZnT7, ZnT9, ZIP3, ZIP5, B 0 AT1, LAT1, y+LAT2, rBAT and PepT1 in the jejunum of broilers at 28 (A) and 39 d (B) of age. ZnT1, zinc transporter 1; ZnT4, zinc transporter 4; ZnT5, zinc transporter 5; ZnT7, zinc transporter 7; ZnT9, zinc transporter 9; ZIP3, Zrt-irt-like protein 3; ZIP5, Zrt-irt-like protein 5; B 0 AT1, B-0-system neutral amino acid co-transporter; LAT1, L-type amino acid transporter 1; y+LAT2, y+L-type amino acid transporter 2; rBAT, b 0 , + -type amino acid transporter; PepT1, peptide-transporter 1; ZnS, Zn sulfate; Zn-Prot M, Zn proteinate with moderate chelation strength (Q f = 51.6).

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Western Blot

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the protein expression levels of Zn, amino acid and small peptide transporters in the ileum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Representative immunoblots of ZnT1, ZnT4, ZnT5, ZnT7, ZnT9, ZIP3, ZIP5, B 0 AT1, LAT1, y+LAT2, rBAT and PepT1 in the ileum of broilers at 28 (A) and 39 d (B) of age. ZnT1, zinc transporter 1; ZnT4, zinc transporter 4; ZnT5, zinc transporter 5; ZnT7, zinc transporter 7; ZnT9, zinc transporter 9; ZIP3, Zrt-irt-like protein 3; ZIP5, Zrt-irt-like protein 5; B 0 AT1, B-0-system neutral amino acid co-transporter; LAT1, L-type amino acid transporter 1; y+LAT2, y+L-type amino acid transporter 2; rBAT, b 0 , + -type amino acid transporter; PepT1, peptide-transporter 1; ZnS, Zn sulfate; Zn-Prot M, Zn proteinate with moderate chelation strength (Q f = 51.6).

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Western Blot

Journal: Nature Communications

Article Title: Physiological and therapeutic regulation of glucose homeostasis by upper small intestinal PepT1-mediated protein sensing

doi: 10.1038/s41467-018-03490-8

Figure Lengend Snippet: Inhibition of peptide transporter 1 (PepT1) reverses the ability of casein infusion to increase glucose tolerance in healthy rodents. Percentage of change in plasma glucose levels ( a ), integrated area under the curve (AUC, b ), and plasma insulin levels ( c ) over time during the IVGTT for healthy rats that received an upper small intestinal infusion of saline ( n = 18), casein ( n = 24), saline+the competitive PepT1 antagonist 4-aminomethylbenzoic acid (4-AMBA, n = 7), or casein+4-AMBA ( n = 6). Values are presented as mean ± s.e.m., where asterisk (*) represents p < 0.05 compared to saline control and hash ( # ) represents p < 0.05 compared to casein+4-AMBA (assessed using ANOVA with Tukey post-hoc test)

Article Snippet: A subset of rats received upper small intestinal injections of purified lentiviral particles expressing mismatch or rat PepT1 shRNA (1 × 10 6 infection units, Santa Cruz, Dallas, TX, USA), which has previously been optimized to specifically infect the upper and not the lower small intestine .

Techniques: Inhibition, Clinical Proteomics, Saline, Control

Journal: Nature Communications

Article Title: Physiological and therapeutic regulation of glucose homeostasis by upper small intestinal PepT1-mediated protein sensing

doi: 10.1038/s41467-018-03490-8

Figure Lengend Snippet: Upper small intestinal casein infusion lowers glucose production through activation of PepT1 in healthy rodents. In vivo glucose kinetics were assessed in conscious, unrestrained healthy rats using the pancreatic (basal insulin) euglycemic clamp as outlined in a . Rates of glucose infusion ( b ), glucose production (GP, c ), and glucose uptake ( d ) in rats that received an upper small intestinal (S.I). infusion of saline ( n = 10) or casein ( n = 9), intravenous (IV) infusion of casein ( n = 5), upper small intestinal infusion of saline+4-AMBA ( n = 6), casein+4-APAA ( n = 5), or casein+4-AMBA ( n = 6). Relative PepT1 mRNA expression in the mucosal layer isolated from S.I. segments ~6–10 cm (upper S.I., e ) or ~25–30 cm (mid-S.I., f ) distal to the pyloric sphincter in rats that received an upper S.I. lentiviral infection with control mismatch ( n = 6) or PepT1 shRNA ( n = 6) particles. Rates of glucose production ( g ) and glucose infusion ( h ) in rats that received mismatch shRNA lentiviral infection+saline infusion ( n = 6), mismatch+casein infusion ( n = 6), PepT1 shRNA lentiviral infection+saline infusion ( n = 5), or PepT1 shRNA+casein infusion ( n = 6). Rates of glucose infusion ( i ) and glucose production ( j ) in rats that received saline+the GLP-1 receptor antagonist exendin-9 ( n = 3) or casein+exendin-9 ( n = 6). Values are presented as mean ± s.e.m., where basal represents the average GP of t = 60–90, clamp represents the average GP of t = 190–200 and asterisk (*) represents p < 0.05 compared to saline control. Statistical significance was determined using an unpaired, two-tailed t -test (two groups) or ANOVA with Tukey post-hoc test (3+ groups)

Article Snippet: A subset of rats received upper small intestinal injections of purified lentiviral particles expressing mismatch or rat PepT1 shRNA (1 × 10 6 infection units, Santa Cruz, Dallas, TX, USA), which has previously been optimized to specifically infect the upper and not the lower small intestine .

Techniques: Activation Assay, In Vivo, Saline, Expressing, Isolation, Infection, Control, shRNA, Two Tailed Test

Journal: Nature Communications

Article Title: Physiological and therapeutic regulation of glucose homeostasis by upper small intestinal PepT1-mediated protein sensing

doi: 10.1038/s41467-018-03490-8

Figure Lengend Snippet: Upper small intestinal PepT1-mediated protein sensing is physiologically relevant during refeeding in healthy rats. Conscious, unrestrained healthy rats underwent a fasting–refeeding protocol in which a low protein (LP) or casein-enriched high protein (HP) diet was offered ad libitum as outlined in a . Percentage of change in plasma glucose levels and integrated area under the curve ( b , c ), cumulative food intake ( d ), and plasma insulin levels ( e ) were monitored in rats that received LP or HP chow with an upper small intestinal (S.I.) infusion of saline (LP: n = 7, HP: n = 9) or 4-AMBA (LP: n = 6, HP: n = 9). Values are presented as mean ± s.e.m., where asterisk (*) represents p < 0.05 compared to LP saline and hash ( # ) represents p < 0.05 compared to HP+4-AMBA. For the analysis of a single time point, ANOVA with Tukey post-hoc test was used to determine statistical significance

Article Snippet: A subset of rats received upper small intestinal injections of purified lentiviral particles expressing mismatch or rat PepT1 shRNA (1 × 10 6 infection units, Santa Cruz, Dallas, TX, USA), which has previously been optimized to specifically infect the upper and not the lower small intestine .

Techniques: Clinical Proteomics, Saline

Journal: Nature communications

Article Title: Physiological and therapeutic regulation of glucose homeostasis by upper small intestinal PepT1-mediated protein sensing.

doi: 10.1038/s41467-018-03490-8

Figure Lengend Snippet: Fig. 3 Upper small intestinal casein infusion lowers glucose production through activation of PepT1 in healthy rodents. In vivo glucose kinetics were assessed in conscious, unrestrained healthy rats using the pancreatic (basal insulin) euglycemic clamp as outlined in a. Rates of glucose infusion (b), glucose production (GP, c), and glucose uptake (d) in rats that received an upper small intestinal (S.I). infusion of saline (n = 10) or casein (n = 9), intravenous (IV) infusion of casein (n = 5), upper small intestinal infusion of saline+4-AMBA (n = 6), casein+4-APAA (n = 5), or casein+4-AMBA (n = 6). Relative PepT1 mRNA expression in the mucosal layer isolated from S.I. segments ~6–10 cm (upper S.I., e) or ~25–30 cm (mid-S.I., f) distal to the pyloric sphincter in rats that received an upper S.I. lentiviral infection with control mismatch (n = 6) or PepT1 shRNA (n = 6) particles. Rates of glucose production (g) and glucose infusion (h) in rats that received mismatch shRNA lentiviral infection+saline infusion (n = 6), mismatch+casein infusion (n = 6), PepT1 shRNA lentiviral infection+saline infusion (n = 5), or PepT1 shRNA+casein infusion (n = 6). Rates of glucose infusion (i) and glucose production (j) in rats that received saline+the GLP-1 receptor antagonist exendin-9 (n = 3) or casein+exendin-9 (n = 6). Values are presented as mean ± s.e.m., where basal represents the average GP of t = 60–90, clamp represents the average GP of t = 190–200 and asterisk (*) represents p < 0.05 compared to saline control. Statistical significance was determined using an unpaired, two-tailed t-test (two groups) or ANOVA with Tukey post-hoc test (3+ groups)

Article Snippet: A subset of rats received upper small intestinal injections of purified lentiviral particles expressing mismatch or rat PepT1 shRNA (1 × 106 infection units, Santa Cruz, Dallas, TX, USA), which has previously been optimized to specifically infect the upper and not the lower small intestine42.

Techniques: Activation Assay, In Vivo, Saline, Expressing, Isolation, Infection, Control, shRNA, Two Tailed Test

Journal: Frontiers in Endocrinology

Article Title: The Dipeptide Pro-Gly Promotes IGF-1 Expression and Secretion in HepG2 and Female Mice via PepT1-JAK2/STAT5 Pathway

doi: 10.3389/fendo.2018.00424

Figure Lengend Snippet: The primer sequences used for real-time quantitative PCR.

Article Snippet: The HepG2 cells were transfected with 4 pmol of siRNA specific for PepT1 (GenePharma Co., Ltd, Shanghai, China) or scrambled siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for 6 h according to the manufacturer's instructions.

Techniques: Amplification

Journal: Frontiers in Endocrinology

Article Title: The Dipeptide Pro-Gly Promotes IGF-1 Expression and Secretion in HepG2 and Female Mice via PepT1-JAK2/STAT5 Pathway

doi: 10.3389/fendo.2018.00424

Figure Lengend Snippet: Knockdown of PepT1 eliminated the increase of IGF-1 expression induced by Pro-Gly in the HepG2 cells. (A) Relative PepT1 mRNA level in HepG2 cells after 24 h incubation in the presence of Pro-Gly and/or PepT1 siRNA (si PePT1). GAPDH was used as the housekeeping gene. (B) The relative PepT1 mRNA level in response to PepT1 siRNA in HepG2 cells after 6 h incubation. GAPDH was used as the housekeeping gene. (C) Western blot analysis of prepro IGF-1 in the HepG2 cells after 24 h incubation in the presence of Pro-Gly (0.5 mM) and/or PepT1 siRNA. β-actin was used as loading control. The panels shown are the representative bands of 3 independent experiments with 6 replicates. (D) Mean ± SEM of immunoblotting bands of prepro IGF-1 ( n = 6). Bars that do not share the same letter are significantly different ( P < 0.05).

Article Snippet: The HepG2 cells were transfected with 4 pmol of siRNA specific for PepT1 (GenePharma Co., Ltd, Shanghai, China) or scrambled siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for 6 h according to the manufacturer's instructions.

Techniques: Knockdown, Expressing, Incubation, Western Blot, Control

Journal: Frontiers in Endocrinology

Article Title: The Dipeptide Pro-Gly Promotes IGF-1 Expression and Secretion in HepG2 and Female Mice via PepT1-JAK2/STAT5 Pathway

doi: 10.3389/fendo.2018.00424

Figure Lengend Snippet: Pro-Gly activated JAK2/STAT5 signaling pathway in a PepT1-dependent manner. (A) Western blot analysis of phospho-JAK2 (p-JAK2), JAK2, phospho-STAT5 (p-STAT5), and STAT5 in HepG2 cells after 24 h incubation in the presence of Pro-Gly (0.5 mM) and/or PepT1 siRNA. β-actin was used as loading control. The panels shown are the representative bands of 3 independent experiments with 6 replicates. (B) Mean ± SEM of immunoblotting bands of p-JAK2/JAK2 and p-STAT5/STAT5 ( n = 6). The intensities of the bands were expressed as the arbitrary units. Bars that do not share the same letter are significantly different ( P < 0.05). (C) Interaction (binding) between JAK2 and STAT5 detected by co-IP. HepG2 cells were exposed to 0.5 mM Pro-Gly or 0.5 mM Pro+Gly for 24 h. (D) HepG2 cells were incubated in the presence of Pro-Gly (0.5 mM) and/or AZD1480 (1 μM) for 6 h and phospho-STAT5 translocation to nuclei was detected by ICC. Scale bar, 10 μm. The IP and ICC experiments were conducted independently for 3 times, with 3 replicates each time.

Article Snippet: The HepG2 cells were transfected with 4 pmol of siRNA specific for PepT1 (GenePharma Co., Ltd, Shanghai, China) or scrambled siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for 6 h according to the manufacturer's instructions.

Techniques: Western Blot, Incubation, Control, Binding Assay, Co-Immunoprecipitation Assay, Translocation Assay

Journal: Frontiers in Endocrinology

Article Title: The Dipeptide Pro-Gly Promotes IGF-1 Expression and Secretion in HepG2 and Female Mice via PepT1-JAK2/STAT5 Pathway

doi: 10.3389/fendo.2018.00424

Figure Lengend Snippet: Acute or chronic injection of Pro-Gly, but not Pro plus Gly (Pro+Gly), stimulated IGF-1 expression and secretion in mice. (A–D) The 18 6-week-old female mice were intraperitoneal injected with physiological saline (Control, n = 6), Pro-Gly (100 mg/kg, n = 6), or Pro (58 mg/kg) plus Gly (38 mg/kg) (Pro+Gly, n = 6) for 1 h. Effects of acute injection of Pro-Gly or Pro+Gly on IGF-1 mRNA level (A) , prepro IGF-1 protein expression and activation of JAK2/STAT5 signaling pathway (B,C) in mice liver and serum level of IGF-1 (D) . β-actin was used as loading control. The intensities of the bands were expressed as the arbitrary units. (E–H) The 30 4-week-old female mice were intraperitoneal injected with physiological saline (Control, n = 10), Pro-Gly (150 mg/kg, n = 10), or Pro (87 mg/kg) plus Gly (57 mg/kg) (Pro+Gly, n = 10) every other day for 35 days. Effects of chronic injection of Pro-Gly or Pro+Gly on PepT1 mRNA level (E) , prepro IGF-1 protein expression and activation of JAK2/STAT5 signaling pathway (F,G) in mice liver and serum level of IGF-1 (H) . β-actin was used as loading control. The intensities of the bands were expressed as the arbitrary units. * P < 0.05. Bars that do not share the same letter are significantly different ( P < 0.05).

Article Snippet: The HepG2 cells were transfected with 4 pmol of siRNA specific for PepT1 (GenePharma Co., Ltd, Shanghai, China) or scrambled siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for 6 h according to the manufacturer's instructions.

Techniques: Injection, Expressing, Saline, Control, Activation Assay