Journal: Annals of Translational Medicine

Article Title: Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation

doi: 10.21037/atm-21-2829

Figure Lengend Snippet: Primers used for real-time quantitative PCR

Article Snippet: Detailed information of all antibodies is given in . table ft1 table-wrap mode="anchored" t5 caption a7 Marker (species) Application (dilution) Distributor (catalog number) Primary antibodies FoxA2 (rabbit) IF (1:200) Proteintech (22474-1-AP) Sox17 (mouse) IF (1:200) R&D (MAB1924-SP) AFP (rabbit) IF (1:100) Proteintech (14550-1-AP) HNF4α (mouse) IF (1:500) Santa Cruz (sc-374229) ALB (rabbit) IF (1:200) Proteintech (16475-1-AP) A1AT (mouse) IF (1:100) Proteintech (66135-1-Ig) ALB (rabbit) WB (1:5,000) Proteintech (16475-1-AP) AFP (rabbit) WB (1:1,000) Santa Cruz (sc-8399) A1AT (mouse) WB (1:5,000) Proteintech (66135-1-Ig) HNF4α (rabbit) WB (1:1,000) ACTIVE MOTIF (61190) NTCP (mouse) WB (1:1,000) Santa Cruz (Sc-518115) FXR (mouse) WB (1:1,000) Santa Cruz (SC-25309) GAPDH (rabbit) WB (1:5,000) Proteintech (10494-1-AP) Secondary antibodies Goat anti-rabbit IgG-HRP antibody WB (1:10,000) Boster (BA1051) Goat anti- mouse IgG-HRP antibody WB (1:10,000) Boster (BA1054) Goat anti-rabbit IgG secondary antibody IF (1:100) Boster (BA1032) Goat anti-mouse IgG secondary antibody IF (1:100) Boster (BA1101) Open in a separate window caption a8 Antibodies used for immunofluorescence and western blotting

Techniques: Sequencing

Journal: Annals of Translational Medicine

Article Title: Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation

doi: 10.21037/atm-21-2829

Figure Lengend Snippet: Antibodies used for immunofluorescence and western blotting

Article Snippet: Detailed information of all antibodies is given in . table ft1 table-wrap mode="anchored" t5 caption a7 Marker (species) Application (dilution) Distributor (catalog number) Primary antibodies FoxA2 (rabbit) IF (1:200) Proteintech (22474-1-AP) Sox17 (mouse) IF (1:200) R&D (MAB1924-SP) AFP (rabbit) IF (1:100) Proteintech (14550-1-AP) HNF4α (mouse) IF (1:500) Santa Cruz (sc-374229) ALB (rabbit) IF (1:200) Proteintech (16475-1-AP) A1AT (mouse) IF (1:100) Proteintech (66135-1-Ig) ALB (rabbit) WB (1:5,000) Proteintech (16475-1-AP) AFP (rabbit) WB (1:1,000) Santa Cruz (sc-8399) A1AT (mouse) WB (1:5,000) Proteintech (66135-1-Ig) HNF4α (rabbit) WB (1:1,000) ACTIVE MOTIF (61190) NTCP (mouse) WB (1:1,000) Santa Cruz (Sc-518115) FXR (mouse) WB (1:1,000) Santa Cruz (SC-25309) GAPDH (rabbit) WB (1:5,000) Proteintech (10494-1-AP) Secondary antibodies Goat anti-rabbit IgG-HRP antibody WB (1:10,000) Boster (BA1051) Goat anti- mouse IgG-HRP antibody WB (1:10,000) Boster (BA1054) Goat anti-rabbit IgG secondary antibody IF (1:100) Boster (BA1032) Goat anti-mouse IgG secondary antibody IF (1:100) Boster (BA1101) Open in a separate window caption a8 Antibodies used for immunofluorescence and western blotting

Techniques: Immunofluorescence, Western Blot

Journal: Annals of Translational Medicine

Article Title: Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation

doi: 10.21037/atm-21-2829

Figure Lengend Snippet: Small molecules efficiently induce DE to differentiate to hepatic progenitors. (A) Morphological changes during hepatic progenitor differentiation. Scale bar =100 µM. (B) qRT-PCR analysis of the mRNA level of hepatic progenitor specific markers expressed in small molecule or growth-factor groups. (C) Immunofluorescent of AFP and HNF4α protein level expression at day 5 (small molecule-induced hepatic progenitor) and day 10 (growth-factor hepatic progenitor). Scale bar =100 µM. (D) Percentage of AFP+ HNF4α+ in immunofluorescence of hepatic progenitors. Undifferentiated human MSCs used as control. All data presented as the mean of at least three independent experiments. *P value <0.05. qRT-PCR, quantitative real-time PCR; SM, small molecule; GF, growth factor; MSC, mesenchymal stem cell; DE, definitive endoderm.

Article Snippet: Detailed information of all antibodies is given in . table ft1 table-wrap mode="anchored" t5 caption a7 Marker (species) Application (dilution) Distributor (catalog number) Primary antibodies FoxA2 (rabbit) IF (1:200) Proteintech (22474-1-AP) Sox17 (mouse) IF (1:200) R&D (MAB1924-SP) AFP (rabbit) IF (1:100) Proteintech (14550-1-AP) HNF4α (mouse) IF (1:500) Santa Cruz (sc-374229) ALB (rabbit) IF (1:200) Proteintech (16475-1-AP) A1AT (mouse) IF (1:100) Proteintech (66135-1-Ig) ALB (rabbit) WB (1:5,000) Proteintech (16475-1-AP) AFP (rabbit) WB (1:1,000) Santa Cruz (sc-8399) A1AT (mouse) WB (1:5,000) Proteintech (66135-1-Ig) HNF4α (rabbit) WB (1:1,000) ACTIVE MOTIF (61190) NTCP (mouse) WB (1:1,000) Santa Cruz (Sc-518115) FXR (mouse) WB (1:1,000) Santa Cruz (SC-25309) GAPDH (rabbit) WB (1:5,000) Proteintech (10494-1-AP) Secondary antibodies Goat anti-rabbit IgG-HRP antibody WB (1:10,000) Boster (BA1051) Goat anti- mouse IgG-HRP antibody WB (1:10,000) Boster (BA1054) Goat anti-rabbit IgG secondary antibody IF (1:100) Boster (BA1032) Goat anti-mouse IgG secondary antibody IF (1:100) Boster (BA1101) Open in a separate window caption a8 Antibodies used for immunofluorescence and western blotting

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Control, Real-time Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation

doi: 10.21037/atm-21-2829

Figure Lengend Snippet: Characterization of small molecule- and growth factor-iHeps. (A) Glycogen storage analysis via PAS staining in small molecule- and growth factor-iHeps. Undifferentiated MSCs used as the control. (B) ICG intake, (C) LDL uptake, scale bar =100 µM. (D) Urea production, (E) Western blots for ALB, A1AT, HNF4α, AFP, NTXP, and FXR expression. GAPDH used as a control. (F) mRNA levels of CYP genes were determined by qRT-PCR in human hUC-MSC-iHeps. All data are presented as the mean of at least three independent experiments. *P value <0.05; **P value <0.01. qRT-PCR, Quantitative real-time PCR; MSC, mesenchymal stem cell; GF, growth factor; SM, small molecule; HGF, hepatocyte growth factor; iHeps, induced hepatocytes; FH1, functional hit 1.

Article Snippet: Detailed information of all antibodies is given in . table ft1 table-wrap mode="anchored" t5 caption a7 Marker (species) Application (dilution) Distributor (catalog number) Primary antibodies FoxA2 (rabbit) IF (1:200) Proteintech (22474-1-AP) Sox17 (mouse) IF (1:200) R&D (MAB1924-SP) AFP (rabbit) IF (1:100) Proteintech (14550-1-AP) HNF4α (mouse) IF (1:500) Santa Cruz (sc-374229) ALB (rabbit) IF (1:200) Proteintech (16475-1-AP) A1AT (mouse) IF (1:100) Proteintech (66135-1-Ig) ALB (rabbit) WB (1:5,000) Proteintech (16475-1-AP) AFP (rabbit) WB (1:1,000) Santa Cruz (sc-8399) A1AT (mouse) WB (1:5,000) Proteintech (66135-1-Ig) HNF4α (rabbit) WB (1:1,000) ACTIVE MOTIF (61190) NTCP (mouse) WB (1:1,000) Santa Cruz (Sc-518115) FXR (mouse) WB (1:1,000) Santa Cruz (SC-25309) GAPDH (rabbit) WB (1:5,000) Proteintech (10494-1-AP) Secondary antibodies Goat anti-rabbit IgG-HRP antibody WB (1:10,000) Boster (BA1051) Goat anti- mouse IgG-HRP antibody WB (1:10,000) Boster (BA1054) Goat anti-rabbit IgG secondary antibody IF (1:100) Boster (BA1032) Goat anti-mouse IgG secondary antibody IF (1:100) Boster (BA1101) Open in a separate window caption a8 Antibodies used for immunofluorescence and western blotting

Techniques: Staining, Control, Western Blot, Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Functional Assay

Journal: Annals of Translational Medicine

Article Title: Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation

doi: 10.21037/atm-21-2829

Figure Lengend Snippet: Use of FH1-based differentiation protocol with human iPSC-derived MSCs. (A) Representative cell morphology of hepatocyte-like cells induced from human iMSCs. Scale bar =100 µM. (B) Immunofluorescence of ALB and A1AT protein expression levels at day 10 in human iMSCs-derived hepatocytes. Scale bar =100 µM. (C) Percentage of ALB + A1AT + in immunofluorescence of hepatocyte cells from iMSC (D) qRT-PCR of hepatocyte-specific markers expressed in iMSCs-derived hepatocytes. Undifferentiated human MSCs used as control. (E) ICG intake. (F) LDL uptake. (G) Glycogen storage analysis via PAS staining in iMSC-derived hepatocytes. Scale bar =100 µM. (H) Western blots for ALB, A1AT, HNF4α, NTXP, and FXR expression. GAPDH used as a control. All data are presented as the mean of at least three independent experiments. qRT-PCR, Quantitative real-time PCR; MSC, mesenchymal stem cell; iMSC, induced pluripotent MSCs; iHeps, induced hepatocytes; FH1, functional hit 1.

Article Snippet: Detailed information of all antibodies is given in . table ft1 table-wrap mode="anchored" t5 caption a7 Marker (species) Application (dilution) Distributor (catalog number) Primary antibodies FoxA2 (rabbit) IF (1:200) Proteintech (22474-1-AP) Sox17 (mouse) IF (1:200) R&D (MAB1924-SP) AFP (rabbit) IF (1:100) Proteintech (14550-1-AP) HNF4α (mouse) IF (1:500) Santa Cruz (sc-374229) ALB (rabbit) IF (1:200) Proteintech (16475-1-AP) A1AT (mouse) IF (1:100) Proteintech (66135-1-Ig) ALB (rabbit) WB (1:5,000) Proteintech (16475-1-AP) AFP (rabbit) WB (1:1,000) Santa Cruz (sc-8399) A1AT (mouse) WB (1:5,000) Proteintech (66135-1-Ig) HNF4α (rabbit) WB (1:1,000) ACTIVE MOTIF (61190) NTCP (mouse) WB (1:1,000) Santa Cruz (Sc-518115) FXR (mouse) WB (1:1,000) Santa Cruz (SC-25309) GAPDH (rabbit) WB (1:5,000) Proteintech (10494-1-AP) Secondary antibodies Goat anti-rabbit IgG-HRP antibody WB (1:10,000) Boster (BA1051) Goat anti- mouse IgG-HRP antibody WB (1:10,000) Boster (BA1054) Goat anti-rabbit IgG secondary antibody IF (1:100) Boster (BA1032) Goat anti-mouse IgG secondary antibody IF (1:100) Boster (BA1101) Open in a separate window caption a8 Antibodies used for immunofluorescence and western blotting

Techniques: Derivative Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Control, Staining, Western Blot, Real-time Polymerase Chain Reaction, Functional Assay

Journal: Cancers

Article Title: Partial Hepatectomy Promotes the Development of KRASG12V-Induced Hepatocellular Carcinoma in Zebrafish

doi: 10.3390/cancers16101793

Figure Lengend Snippet: Effects of PH on the progress of HCC-associated characteristics in kras+ zebrafish. ( A ) IF staining of PCNA and HNF4α in the PH/sham surgery-treated kras+ zebrafish liver 3 days after Dox induction. ( B ) Quantification of PCNA and HNF4α-double positive proliferating hepatocytes based on ( A ) ( n = 4). Circles indicate values of individual samples. ( C – E ) Expression of ( C ) ccnb1 and cdk1, ( D ) col1a1b, lama5, ( E ) acta2 and tgfb1b in the PH/sham surgery-treated kras+ zebrafish livers 3 days after Dox induction as determined by RT-qPCR ( n = 4). ( F ) Expression of kras G12V in the PH/sham surgery-treated kras+ zebrafish livers 3 days after Dox induction as determined by RT-qPCR ( n = 3). Scale Bar = 20 μm. ns p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: The antibodies used in this study and their dilution ratio are Mouse anti-PCNA (sc-56; Santa Cruz) (1:200), Rabbit anti-HNF4α (ab201460; Abcam, Cambridge, UK) (1:2000), Rabbit anti-HIF1α (ab114977; Abcam) (1:200), Goat anti-rabbit IgG Alexa Fluor™ 488 (A11034; Invitrogen) (1:500), and Goat anti-mouse IgG Alexa Fluor™ 568 (A11031; Invitrogen) (1:500).

Techniques: Staining, Expressing, Quantitative RT-PCR

Journal: Cell

Article Title: Tissue Repair Signals and In Vitro Culture: Inflammatory Cytokine TNFα Promotes the Expansion of Primary Hepatocytes in 3D Culture

doi: 10.1016/j.cell.2018.11.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Polyclonal anti- Hnf4α (Clone H-171) , Santa Cruz Biotechnology , Cat#SC-8987; RRID:AB_2116913.

Techniques: Virus, Recombinant, RNAscope, Reverse Transcription, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Expressing, Plasmid Preparation, Software, Cell Analysis

Journal: Cell

Article Title: Tissue Repair Signals and In Vitro Culture: Inflammatory Cytokine TNFα Promotes the Expansion of Primary Hepatocytes in 3D Culture

doi: 10.1016/j.cell.2018.11.012

Figure Lengend Snippet: Antibodies

Article Snippet: Rabbit Polyclonal anti- Hnf4α (Clone H-171) , Santa Cruz Biotechnology , Cat#SC-8987; RRID:AB_2116913.

Techniques: Produced