Journal: Hepatology Communications

Article Title: Arrb2 in hepatocytes promotes M2 macrophage polarization, ameliorates hepatic ischemia–reperfusion injury through upregulating metabolite 6-ketoLCA

doi: 10.1097/HC9.0000000000000916

Figure Lengend Snippet: 6-ketoLCA upregulated by Arrb2 in hepatocytes promotes M2 macrophage polarization, alleviating hepatic IRI. (A–E) Liver and serum samples from liver-specific Arrb2-CKO and WT mice with or without 6-ketoLCA treatment after I/R (n=6). (A) HE and TUNEL staining were analyzed microscopically. Scale bar: 200 μm; Suzuki score and necrosis area were examined. (B) Quantitative analysis of the bile acid metabolite 6-ketoLCA in liver tissue. (C) Liver function levels (ALT, AST, TBA, GGT) were analyzed between groups. (D) qRT-PCR analysis of mouse liver tissue mRNA expression for inflammatory factors (IL-6, TNF-α, IL-10, TGF-β). (E) iNOS, CD86, CD206, and CD163 expression in PMMs was assessed by WB analysis. (F) The CCK8 assay measured the viability of RAW264.7 cells exposed to varying concentrations and durations of 6-ketoLCA (n=3). (G, I) Co-culture of RAW264.7 and AML12 cells. (G) ELISA for culture medium supernatant inflammatory factors (IL-6, TNF-α, IL-10, TGF-β). (I) BCL2, BAX, and GAPDH expression in AML12 cells was assessed by WB analysis. (H, J) Flow cytometry detected the proportions of M1 (CD86, PE) and M2 (CD206, APC) macrophages treated with IL-4 and 6-ketoLCA. The ratio of M1/M2 macrophages was measured. All results are presented as mean ± SD, and statistical significance was assessed. * p <0.05, ** p <0.01, and *** p <0.001. Abbreviations: ALT, alanine transaminase; AST, aspartate transaminase; CKO, conditional knockout; GGT, gamma-glutamyl transferase; HE, hematoxylin and eosin; I/R, ischemia/reperfusion; IL-6, interleukin 6; IL-10, interleukin 10; IRI, ischemia–reperfusion injury; qRT-PCR, quantitative reverse transcription polymerase chain reaction; PMMs, primary mouse macrophages; TBA, total bile acids; TGF-β, transforming growth factor-β; TNF-α, tumor necrosis factor-α; TUNEL, terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling; WB, western blotting; WT, wild type.

Article Snippet: RAW264.7 were cultured in 10% FBS RPMI-1640 medium with 6-ketoLCA (100 ng/mL) and IL-4 (20 ng/mL) to induce polarization to M2-like macrophage. shRNA targeting TGR5 (GeneCopoeia, Rockville, MD, USA) was transfected into RAW264.7 cells utilizing Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer’s protocol to achieve TGR5 knockdown.

Techniques: TUNEL Assay, Staining, Quantitative RT-PCR, Expressing, CCK-8 Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Knock-Out, Reverse Transcription, Polymerase Chain Reaction, End Labeling, Western Blot

Journal: Molecules and Cells

Article Title: Deficient chaperone-mediated autophagy in macrophages aggravates colitis and colitis-associated tumorigenesis in mice

doi: 10.1016/j.mocell.2025.100298

Figure Lengend Snippet: The expression of LAMP2A in CRC tissues. (A) Immunofluorescence analysis of LAMP2A (red) and the macrophage marker CD68 (green) in human CRC tissues and paracancerous colitis tissues. n = 6 per group. Scale bar = 50 µm. (B) Quantification of CD68 + LAMP2A + cells' human CRC tissues and paracancerous colitis tissues. n = 6 per group. Statistical analysis was conducted using an unpaired Student’s t -test. Data are expressed as mean ± SD. (C) Immunofluorescence analysis of LAMP2A (red) and the macrophage marker CD68 (green) in colon paraffin sections from mouse models of colitis and CRC. n = 6 per group. Scale bar = 50 µm. (D) Quantification of CD68 + LAMP2A + cells in mouse colitis and CRC tissues. n = 6 per group. Statistical analysis was conducted using an unpaired Student’s t -test. Data are expressed as mean ± SD. (E) RT-PCR analysis of LAMP2A mRNA expression in macrophages isolated from mouse colitis and CRC tissues. The y-axis represents the relative fold change in LAMP2A mRNA expression, normalized to the housekeeping gene β-actin and calculated using the ΔΔCt method. The colorectum tissues from three mice were pooled as one sample for macrophage isolation. n = 5 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (F-G) Representative western blot analysis and quantification of LAMP2A and HSC70 in macrophages isolated from mouse colitis and CRC tissues. Macrophages were isolated from colorectum tissues by the magnetic bead sorting method. The colorectum tissues from three mice were pooled as one sample for macrophage isolation. n = 5 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test.

Article Snippet: The primary antibodies used in this study include LAMP2A (ab125068, Abcam), PCNA ( OM267735 , Omnimabs), Hif-1α (ab179483, Abcam), VEGFR2 (2479, CST), IL-1β (ab9722, Abcam), Pro-IL-1β (#12426, CST), and β-actin (#4970, CST).

Techniques: Expressing, Immunofluorescence, Marker, Reverse Transcription Polymerase Chain Reaction, Isolation, Western Blot

Journal: Molecules and Cells

Article Title: Deficient chaperone-mediated autophagy in macrophages aggravates colitis and colitis-associated tumorigenesis in mice

doi: 10.1016/j.mocell.2025.100298

Figure Lengend Snippet: Effects of LAMP2A knockout and knock-in in macrophages on colitis in C57BL/6 mice. (A) Body weight (% of Baseline), (B) macroscopic appearances, (C) colon lengths, and (D) disease activity index (DAI) scores of LAMP2A-mØKO mice and their littermate controls (WT) administered with regular water (H 2 O) or DSS for 7 days. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis of body weight (% of baseline) was conducted using a mixed-effects model followed by Sidak’s multiple comparisons test. Statistical analysis in B-D was conducted using one-way ANOVA. (E) H&E staining of colon tissues from LAMP2A-mØKO mice and WT mice administered with H 2 O or DSS for 7 days. The above and below scales are 200 and 20 µm, respectively. n = 6 per group. (F) Immunofluorescence analysis of IL-1β (red) and the macrophage marker CD68 (green) in colon sections from LAMP2A-mØKO mice and WT mice, n = 6 per group. Scale bar = 50 µm. (G) Quantification of CD68 + IL-1β + cells within colitis tissues of LAMP2A-mØKO mice and WT mice administered with H 2 O or DSS for 7 days. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (H-J) Representative western blot analysis and quantification of LAMP2A and IL-1β in whole-colon lysates from LAMP2A-mØKO mice and WT mice. n = 5 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using one-way ANOVA. (K) Body weight (% of Baseline), (L) macroscopic appearances, (M) colon lengths, and (N) disease activity index (DAI) scores of LAMP2A-mØKi mice and their littermate controls (WT) administered with H 2 O or DSS for 7 days. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis of Body weight (% of Student’s t test or one-way ANline) was conducted using a mixed-effects model followed by Sidak’s multiple comparisons test. Statistical analysis in L to N was conducted using one-way ANOVA.

Article Snippet: The primary antibodies used in this study include LAMP2A (ab125068, Abcam), PCNA ( OM267735 , Omnimabs), Hif-1α (ab179483, Abcam), VEGFR2 (2479, CST), IL-1β (ab9722, Abcam), Pro-IL-1β (#12426, CST), and β-actin (#4970, CST).

Techniques: Knock-Out, Knock-In, Activity Assay, Staining, Immunofluorescence, Marker, Western Blot

Journal: Molecules and Cells

Article Title: Deficient chaperone-mediated autophagy in macrophages aggravates colitis and colitis-associated tumorigenesis in mice

doi: 10.1016/j.mocell.2025.100298

Figure Lengend Snippet: Effects of LAMP2A knockout in macrophages on angiogenesis . (A-B) Representative immunohistochemical images and quantification of CD31 in CRC sections from LAMP2A-mØKO mice and WT mice. For CD31+ endothelial cells, quantification was conducted by counting the number of vessels per field of view across 6 randomly selected fields for each sample under ×800 magnifications. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (C-D) Representative immunohistochemical images and quantification of IL-1β in CRC sections from LAMP2A-mØKO mice and WT mice. For IL-1β, the percentage of positive staining area relative to the total tissue area was measured using ImageJ software with a consistent threshold applied to all images. Statistical analysis was conducted using an unpaired Student’s t -test. n = 6 per group. Data are expressed as mean ± SD. (E-H) Representative immunofluorescence analysis and quantification of VEGFA and VEGFR2 in CRC sections from LAMP2A-mØKO mice and WT mice. VEGFA was co-stained with CD68, a marker for macrophages, and VEGFR2 was co-stained with CD31, a marker for endothelial cells. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t-test. Scale bar = 50 µm. (I-L) ELISA analysis of VEGFA and IL-1β in CRC tissue grinding solution and in the cellular supernatant of cultured primary peritoneal macrophages. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (M-N) Wound healing assay to evaluate the migration ability of HUVEC co-cultured with LAMP2A-deficient macrophages or WT macrophages. Five independent experiments were conducted. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (O-P) Tube formation assay to evaluate angiogenesis of HUVEC co-cultured with LAMP2A-deficient macrophages or WT macrophages. HUVECs were treated with IL-1 RI-neutralizing antibody (5 µg/mL) and/or VEGFR2-neutralizing antibody (0.5 µg/mL) for 24 hours. Five independent experiments were conducted. Data are expressed as mean ± SD. Statistical analysis was conducted using one-way ANOVA.

Article Snippet: The primary antibodies used in this study include LAMP2A (ab125068, Abcam), PCNA ( OM267735 , Omnimabs), Hif-1α (ab179483, Abcam), VEGFR2 (2479, CST), IL-1β (ab9722, Abcam), Pro-IL-1β (#12426, CST), and β-actin (#4970, CST).

Techniques: Knock-Out, Immunohistochemical staining, Staining, Software, Immunofluorescence, Marker, Enzyme-linked Immunosorbent Assay, Cell Culture, Wound Healing Assay, Migration, Tube Formation Assay

Journal: Molecules and Cells

Article Title: Deficient chaperone-mediated autophagy in macrophages aggravates colitis and colitis-associated tumorigenesis in mice

doi: 10.1016/j.mocell.2025.100298

Figure Lengend Snippet: LAMP2A deficiency promoted the secretion of IL-1β and VEGFA by upregulating HIF-1α. (A-B) Representative immunohistochemical images and quantitative analysis of HIF-1α expression based on nuclear localization. The graph shows the percentage of cells with positive HIF-1α nuclear staining in CRC sections from LAMP2A-mØKO mice and WT mice. n = 6 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (C-D) Representative western blot analysis and quantification of HIF-1α in macrophages isolated from CRC tissues of LAMP2A-mØKO mice and WT mice. Macrophages were isolated from colorectum tissues by the magnetic bead sorting method. The colorectum tissues from three mice were pooled as one sample for macrophage isolation. n = 5 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (E) RT-PCR analysis of HIF-1α mRNA expression in macrophages isolated from CRC tissues of LAMP2A-mØKO mice and WT mice. The colorectum tissues from 3 mice were pooled as one sample for macrophage isolation. n = 5 per group. Data are expressed as mean ± SD. Statistical analysis was conducted using an unpaired Student’s t -test. (F) Immunofluorescence analysis of LAMP2A (red) and HIF-1α (green) in human CRC tissues. Scale bar = 50 µm. (G) Co-IP assay of the interaction between HIF-1α and LAMP2A in macrophages isolated from human CRC tissues. The tissue lysates immunoprecipitated with either the anti-LAMP2A antibody or rabbit IgG and probed for HIF-1α and LAMP2A. Three independent experiments were conducted. (H-I) ELISA analysis of VEGFA and IL-1β in the cellular supernatant of cultured primary peritoneal macrophages treated with or without lificiguat (YC-1, 1 µM) for 24 hours. Five independent experiments were conducted. Data are expressed as mean ± SD. Statistical analysis was conducted using one-way ANOVA. (J-K) Tube formation assay to evaluate angiogenesis of HUVEC co-cultured with LAMP2A-deficient macrophages or WT macrophages. Macrophages were treated with or without lificiguat (YC-1, 1 µM) for 24 hours. Five independent experiments were conducted. Data are expressed as mean ± SD. Statistical analysis was conducted using one-way ANOVA.

Article Snippet: The primary antibodies used in this study include LAMP2A (ab125068, Abcam), PCNA ( OM267735 , Omnimabs), Hif-1α (ab179483, Abcam), VEGFR2 (2479, CST), IL-1β (ab9722, Abcam), Pro-IL-1β (#12426, CST), and β-actin (#4970, CST).

Techniques: Immunohistochemical staining, Expressing, Staining, Western Blot, Isolation, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Co-Immunoprecipitation Assay, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Cell Culture, Tube Formation Assay

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Representative fluorescence microscopy images of mesenchymal stromal cells (MSCs) infected by IL-4 and/or PDGF-BB lentiviral vectors. GFP-MSCs, MSCs infected with control GFP-positive lentivirus vector; IL4-MSCs, MSCs infected with rIL-4 secreting GFP-positive lentivirus vector; PDGF-BB-MSCs, MSCs infected with hPDGF-BB secreting GFP-positive lentivirus vector; IL4-PDGF-BB-MSCs, previously established IL4-MSCs infected with hPDGF-BB secreting RFP-positive lentivirus vector.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Fluorescence, Microscopy, Infection, Control, Plasmid Preparation

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: The experimental outline of in vitro experiments. GFP-MSCs, IL4-MSCs, PDGF-BB-MSCs, or IL4-PDGF-BB-MSCs were seeded into T75 flasks and cultured in an MSC growth medium for 1 day. For the non-preconditioning MSC groups, the cells were cultured for 3 days with a fresh MSC growth medium. For the preconditioning MSC groups, cells were cultured in the MSC preconditioning medium for 3 days. The cells from all eight groups were washed three times with Dulbecco’s phosphate-buffered saline. The cells were trypsinized and seeded for each experiment, including cell proliferation, IL4 and PDGF-BB expression level, and osteogenic differentiation performed by alkaline phosphatase (ALP) staining and Alizarin red-staining. Abbreviations: MSCs, mesenchymal stromal cells; pMSCs, preconditioning MSCs; TNFα, tumor necrosis factor-alpha; LPS, lipopolysaccharide.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: In Vitro, Cell Culture, Saline, Expressing, Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Fold change in the amount of dsDNA compared to day 1.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques:

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: ( A ) IL4 and PDGF-BB expression levels on day 3 in the IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups. IL4 expression levels on day 3 in the IL4-MSC, IL4-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 77.7 ± 4.4 ng/mL, 34.6 ± 5.7 ng/mL, 16.8 ± 9.4 ng/mL, and 41.8 ± 5.0 ng/mL, respectively. PDGF-BB expression levels on day 3 in the PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 121.7 ± 20.1 pg/mL, 50.3 ± 20.4 pg/mL, 8.7 ± 2.0 pg/mL, 71.5 ± 7.8 pg/mL, respectively. ( B ) IL4 and PDGF-BB expression level-dsDNA ratios on day 3 in the IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups. IL4 expression level-dsDNA ratios on day 3 in the IL4-MSC, IL4-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 7.3 ± 0.6, 1.8 ± 0.2, 1.8 ± 1.0, and 2.3 ± 0.1, respectively. PDGF-BB expression level-dsDNA ratios on day 3 in the PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 5.6 ± 0.9, 3.7 ± 1.4, 1.0 ± 0.3, and 3.9 ± 0.1, respectively.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Expressing

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Alkaline phosphatase-staining in all eight groups 2 weeks after seeding. The percentages of stained area in the GFP-MSC, GFP-pMSC, IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 19.8 ± 5.3%, 28.3 ± 1.4%, 4.3 ± 0.7%, 6.0 ± 2.4%, 44.2 ± 11.7%, 42.5 ± 8.1%, 21.4 ± 1.5%, and 25.7 ± 1.0%, respectively. Abbreviations: ALP, alkaline phosphatase.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: Alizarin red-staining in all eight groups 4 weeks after seeding. The percentages of stained area in the GFP-MSC, GFP-pMSC, IL4-MSC, IL4-pMSC, PDGF-BB-MSC, PDGF-BB-pMSC, IL4-PDGF-BB-MSC, and IL4-PDGF-BB-pMSC groups were 33.1 ± 4.7%, 45.1 ± 4.9%, 0.64 ± 0.08%, 0.58 ± 0.03%, 51.7 ± 14.5%, 40.7 ± 6.7%, 17.3 ± 2.5%, and 41.2 ± 1.7%, respectively.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Staining

Journal: Bioengineering

Article Title: Effect on Osteogenic Differentiation of Genetically Modified IL4 or PDGF-BB Over-Expressing and IL4-PDGF-BB Co-Over-Expressing Bone Marrow-Derived Mesenchymal Stromal Cells In Vitro

doi: 10.3390/bioengineering8110165

Figure Lengend Snippet: The summary of the results.

Article Snippet: The expression levels of IL4 and PDGF-BB on day 3 were measured using the rabbit IL4 ELISA Kit (R&D Systems, Minneapolis, MN, USA) and the human PDGF-BB ELISA kit (R&D Systems, Minneapolis), respectively, according to the manufacturer’s protocols.

Techniques: Expressing

Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison