recombinant murine epidermal growth factor (egf)  (PeproTech)

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: Identification of JH015Y protein. (A) Protein electrophoresis map. (B) Mass spectrum. (C) Analysis of JH015Y protein and its matching degree with protein database data. HIVEP3: HIVEP Zinc Finger 3; A member of the human immunodeficiency virus type 1 enhancer binding protein family. NAC-A/B: domains of NAC transcription factors unique to plants. LIM: A highly conserved double zinc finger domains in evolution.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: Protein Electrophoresis, Virus, Binding Assay

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: Promoting of JH015Y protein on cell proliferation in vitro . (A) Influence of JH015Y protein in the proliferation of HaCaT cells. (B) Influence of JH015Y protein in the proliferation of HSF cells. *** p <0.001, ** p <0.01, * p <0.05. n=5. EGF: Epidermal Growth Factor. HSF: Human Skin Fibroblasts. HaCaT: Human Keratinocytes Cells.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: In Vitro

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: The promoting effect of JH015Y protein on in vitro cell migration. (A) The effect of JH015Y protein on the migration of HaCaT cells. (B) The effect of JH015Y protein on HSF cell migration. (C) The effect of JH015Y protein on cytokine secretion of HaCaT. (D) The effect of JH015Y protein on cytokine secretion in HSF. *** p <0.001, ** p <0.01, * p <0.05. n=5. HSF: Human Skin Fibroblasts. EGF: Epidermal Growth Factor. VEGF: Vascular Endothelial Growth Factor.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: In Vitro, Migration

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: Therapeutic effect of JH015Y protein on acute wounds in vivo . (A) The wound healing rate of rats in different groups and days. (B) Photos of rat wounds from different groups on different days. (C) Masson staining results of skin samples from different groups of rats on different days (scale: 100 μm). **** p <0.0001, *** p <0.001, ** p <0.01, * p <0.05. n=8. EGF: Epidermal Growth Factor.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: In Vivo, Staining

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: Therapeutic effect of JH015Y protein on burn wounds in vivo . (A) Photos of rat wounds from different groups on different days. (B) The healing rate of scalds in rats of different groups and days. (C) Masson staining results of skin samples from different groups of rats on different days (scale: 100 μm). *** p <0.001, ** p <0.01, * p <0.05. n=8. EGF: Epidermal Growth Factor.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: In Vivo, Staining

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: Therapeutic effect of JH015Y protein on diabetic ulcer in vivo . (A) Photos of mouse wounds from different groups on different days. (B) Healing rate of ulcer wounds in mice of different groups and days. (C) Masson staining results of skin samples from different groups of rats on different days (scale: 100 μm). ** p <0.01, * p <0.05. n=8. EGF: Epidermal Growth Factor.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques: In Vivo, Staining

Journal: Biomolecules & Therapeutics

Article Title: An Amphibians-Derived Protein Provides Novel Biotherapeutics for Various Wounds Treatment

doi: 10.4062/biomolther.2024.096

Figure Lengend Snippet: The therapeutic effects of the JH015Y protein in different wounds.

Article Snippet: This study utilized thefollowing listed materials:HaCaTcells (maintained in our laboratory); HSF cells (maintained in our laboratory); JH015Y protein; recombinant murine epidermal growth factor (EGF) (PeproTech, 5 Cedarbrook Drive, Cranbury, NJ, USA); DMEM culture medium; fetal bovine serum (Gibco, Carlsbad, CA, USA); penicillin/streptomycin stock solution (Institute of Biomedical Engineering, Chinese Academy of Medical Sciences, 236 Baidi Road, Tianjin, China); CCK-8 assay kit (Servicebio, Wuhan, China); Clean bench (BSC-1100IIA, Beijing Donglian Haer Instrument Manufacturing Co., Ltd., Beijing, China); CO2 cell culture incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd, Shanghai, China); microplate reader (Jiangnan Corporation, Ningbo, China); Depilatory cream (Veet, London, UK); sterile dressing (6×7 cm, Zhejiang Ou Jie Technology Co., Ltd., Deqing, China); glutaraldehyde (JiZhiSheng Biology, Shanghai, China); OCT embedding agent (Tissue-TEK, Sakura, Torrance, CA, USA); Masson’s trichrome staining kit (Nanjing Kaiji Biotechnology Development Co., Ltd., Nanjing, China); cryostat (CM1950, Leica, Wetzlar, Germany); YLS-5Q desktop constant temperature and pressure scald instrument (Tianjin Norei Xinda Technology Co., Ltd., Tianjin, China); blood glucose meter (Sinocare, Changsha, China); Sprague Dawley rats (male, 120-160 g, four-week-old), C57BLKS/J (db/db) mice (male, 40-50 g, six-week-old), provided by Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China; BCA protein quantification kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); electrophoresis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China); protein molecular weight standards (Shanghai Yaen Biotechnology Co., Ltd., Shanghai, China); sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) gel rapid preparation kit (Dalian Meilun Biotechnology Co., Ltd., Dalian, China); protein loading buffer; chemiluminescence imager (BioRad, Hercules, CA, USA); vertical protein electrophoresis apparatus (Hangzhou Nuoyang Biotechnology Co., Ltd., Hangzhou, China); Maldi TOF mass spectrometer (Bruker, Saarbrucken, Germany); ultra-high resolution mass spectrometer (Thermo Fisher, Waltham, MA, USA).

Techniques:

Journal: bioRxiv

Article Title: Viroporin activity from rotavirus nonstructural protein 4 induces intercellular calcium waves that contribute to pathogenesis

doi: 10.1101/2024.05.07.592929

Figure Lengend Snippet: a) Schematic representation of the recombinant SA11 strains with gene segment 10 encoding NSP4 from either OSUv or OSUa (i). The linearized domain map (ii) illustrates the 3 amino acid differences between OSUv and OSUa NSP4. b ) Quantification of ICWs in MA104-GCaMP6s monolayers infected with mock inoculum, SA11, SA11 expressing gene 10 (NSP4) from OSUv or OSUa, or SA11 expressing the NSP4 from OSUa with one of the 3 amino acids reverted to the OSUv-NSP4 identity in turn. Monolayers were imaged from 8-24 hpi and ICWs quantitated across 3 biological replicates. c ) Growth curves comparing viral yield at 6 time points post-infection. d ) NSP4 detected at 9 hpi in MA104 monolayer lysate following infection at MOI 5 with the indicated strain of recombinant SA11. e ) Representative images of SA11-G10-OSUv and -OSUa infected monolayers expressing ER-localized GFP (green) with NSP4 (magenta) and NSP2 (orange) detected by immunofluorescence at 12 hpi. Scale bar = 10μm. f ) Mander’s coefficient to estimate NSP4 colocalization with Sec61β as a surrogate for ER localization. Combined data from 15 cells across 3 biological replicates. g) Representative intracellular Ca 2+ traces from cells infected with the indicated strain of recombinant SA11. h) Intracellular Ca 2+ spike counts from (i) MA104-GCaMP6s or (ii) LLCMK2 cells (ICW deficient) infected with the indicated strain of recombinant SA11. Combined data from 3 biological replicates. i ) Linear regression estimating the association between intracellular Ca 2+ spike counts in LLCMK2-GCaMP6s cells and ICW counts in MA104 GCaMP6s cells.

Article Snippet: MIOs were maintained in Advanced DMEM/F12 (Gibco) supplemented with 1X GlutaMax (Gibco), 0.01M HEPES, 100 U/ml penicillin-streptomycin (Gibco), 20% Rspo-1 conditioned medium, 5% Noggin conditioned medium, 1mM N2 supplement (Invitrogen), B27 supplement (Invitrogen), 1mM N-acetylcysteine (Sigma-Aldrich), and 50 ng/mL recombinant murine epidermal growth factor (Invitrogen).

Techniques: Recombinant, Infection, Expressing, Immunofluorescence