Journal: World Journal of Stem Cells

Article Title: Patient-derived induced pluripotent stem cells with a MERTK mutation exhibit cell junction abnormalities and aberrant cellular differentiation potential

doi: 10.4252/wjsc.v16.i5.512

Figure Lengend Snippet: Generation and identification of the human induced pluripotent stem cells derived from the retinitis pigmentosa patient. A: Timeline of human induced pluripotent stem cell generation; B: Imaging by phase-contrast microscopy. Scale bar = 200 μm; C: Karyotype analysis of the healthy control (left) and retinitis pigmentosa patient (right); D: Flow cytometry of pluripotency markers SSEA-4 and TRA-1-81; E and F: Immunostaining of pluripotency markers OCT4, NANOG, SOX2, and SSEA4 of the healthy control and retinitis pigmentosa patient. Scale bar = 20 μm; G and H: In vitro differentiation of control (G) and patient (H) induced pluripotent stem cells into three germ layers, endoderm (AFP+), mesoderm (α-SMA+) and ectoderm (GFAP+). Scale bar = 20 μm. PB: Peripheral blood; PBMC: Peripheral blood mononuclear cell; iPSC: Induced pluripotent stem cell.

Article Snippet: Following primary antibodies were used: OCT4 (Cat. #ab18976; Abcam), SOX2 (Cat. #sc-365823; Santa Cruz), NANOG (Cat. #ab80892; Abcam), SSEA4 (Cat. #ab16287; Abcam), GFAP (Cat. #HPA056030; Sigma), α-SMA (Cat. #A5228; Sigma), AFP (Cat. #MAB1368; R&D System).

Techniques: Derivative Assay, Imaging, Microscopy, Control, Flow Cytometry, Immunostaining, In Vitro

Journal: EBioMedicine

Article Title: Tumour-specific activation of a tumour-blood transport improves the diagnostic accuracy of blood tumour markers in mice.

doi: 10.1016/j.ebiom.2024.105178

Figure Lengend Snippet: Fig. 1: Intravenously injected iRGD increases blood AFP levels in HCC-bearing mice. (a) Experimental setup to study the effect of iRGD on blood AFP levels in Huh-7 xenografted nude mice. (b and c) Blood AFP levels before and after intravenous injection of iRGD (b, c, n = 12), RGD control peptide (c (n = 6)), and PBS (c (n = 6)) in mice with HepG2 xenografts. Human AFP was not detectable in the blood of mice without tumours. In (c) the fold increase of AFP with pre-injection level set to 1; lines and error bars represent geometric means and 95% CI. (d) Experimental setup to study the effect of iRGD on blood AFP levels in TGFα/c-myc HCC mice. (e and f) iRGD specifically increased the blood AFP levels in TGFα/c-myc HCC mice. TGFα/c-myc mice (20–24 weeks old) with HCC according to MRI or without HCC (f) were intravenously injected with iRGD (e, f), RGD control peptide (f) or PBS (f). Data are fold changes of blood AFP due to the treatments (n = 48: iRGD; RGD control peptide: n = 34; PBS: n = 15); lines and error bars indicate medians and 95% CI. Dashed line: upper 95% CI increase in blood AFP in the PBS-injected HCC mice. (g) AFP expression in HCCs of TGFα/c-myc mice. HCCs and liver tissues were excised from TGFα/c-myc mice. Pairs of the tissue lysates were analysed for AFP and β-actin content by immunoblotting. Band densities were measured densitometrically. The ratio of AFP/β-actin in the livers was set to 1. (h) iRGD-induced accumulation of Evans blue in HCCs in TGFα/c-myc mice with HCC. TGFα/c-myc mice with HCCs were co- injected with iRGD or PBS (n = 12 per group) and Evans blue (EB). The dye content of the tumours was related to that in the livers; lines and error bars indicate geometric means (b, c, and h) or medians (e and f) and 95% CI; dashed line: upper 95% CI of the measured EB content of a HCC from the PBS-injected animals. (b): paired t test for log-transformed data; (c) One-way ANOVA with multiple comparison post-hoc test for log-transformed data; (e): Wilcoxon signed-rank test for log-transformed data; (f) Kruskal–Wallis test with Dunn’s multiple comparison post- hoc test; (h): two-sample t test. The indicated fold increase in (b, e and h) is the ratio of the geometric means with 95% CI.

Article Snippet: Immunoblotting of lysates obtained from pairs of liver and HCC tissue from TGFα/c-myc mice was performed as described previously.38 Gel-resolved proteins were electrotransferred to nitrocellulose membranes and incubated with antibodies raised against mouse-AFP (#AF5369, R&D Systems, Minneapolis, MN, RRID:AB_2258018) and anti-β-actin (#A2066, Sigma– Aldrich/Merck, RRID:AB_476693).

Techniques: Injection, Control, Expressing, Western Blot, Transformation Assay, Comparison

Journal: EBioMedicine

Article Title: Tumour-specific activation of a tumour-blood transport improves the diagnostic accuracy of blood tumour markers in mice.

doi: 10.1016/j.ebiom.2024.105178

Figure Lengend Snippet: Fig. 3: iRGD-induced elevation of the blood AFP concentration depends on NRP-1 and the tumour blood concentration gradient of AFP. (a) Anti-NRP-1 prevented iRGD-induced increase in the blood AFP concentration. TGFα/c-myc tumour mice that displayed a robust iRGD-induced increase in blood AFP level one week earlier were injected with anti-NRP-1 and the effect of iRGD on blood AFP level was determined (n = 3 per group). Fold increase of AFP with pre-injection level at the first time point was set to 1. Lines and error bars represent medians and 95% CI. (b– e) iRGD-induced elevation of the blood AFP level correlated negatively with the pre-injection blood AFP level in TGFα/c-myc mice (b), DEN- CCl4-HCC mice (c), mice with Huh-7 (d) or HepG2 xenografts (e). Spearman correlation r (b and c) and Pearson correlation r (d and e) with 95% CI, two tailed p-values and the log–log regression lines. (f) iRGD increased the blood AFP levels in HepG2 xenografted nude mice and low basal AFP (<67 ng/ml, n = 36, left panel), but not in animals with high basal AFP (>67 ng/ml, n = 12, right panel). Lines and error bars indicate geometric means and 95% CI. [(g) iRGD increased the blood AFP levels in TGFα/c-myc mice with HCC and normal basal AFP (<67 ng/ml, n = 36, left panel), but not in mice with elevated basal AFP (>67 ng, n = 12, right panel). Lines and error bars represent medians (left) or geometric means (right) with 95% CI. Significance was calculated with one sample t test (a, left) and the unpaired t test (a, right), paired t test (f and g, right) and Wilcoxon matched-pairs signed-rank test (g, left). The indicated fold increase in (f) is the geometric mean ratio with 95% CI. The indicated fold increase in (g) is the median of the ratios with 95% CI.

Article Snippet: Immunoblotting of lysates obtained from pairs of liver and HCC tissue from TGFα/c-myc mice was performed as described previously.38 Gel-resolved proteins were electrotransferred to nitrocellulose membranes and incubated with antibodies raised against mouse-AFP (#AF5369, R&D Systems, Minneapolis, MN, RRID:AB_2258018) and anti-β-actin (#A2066, Sigma– Aldrich/Merck, RRID:AB_476693).

Techniques: Concentration Assay, Injection, Two Tailed Test

Journal: Analytical Chemistry

Article Title: Microarray-Based Multiplexed Scanometric Immunoassay for Protein Cancer Markers Using Gold Nanoparticle Probes

doi: 10.1021/ac9018389

Figure Lengend Snippet: Figure 3. Scanometric identification of three protein cancer markers for eight different samples in buffer after two gold depositions. The concentration of each antigen was 1.4 pM. (1) All targets present; (2) hCG and PSA; (3) hCG and AFP; (4) PSA and AFP; (5) AFP; (6) PSA; (7) hCG; (8) no targets present. The gray scale images from the Verigene Reader system were converted into colored ones using GenePix Pro 6 software (Molecular Devices), and the exposure time was 200 ms.

Article Snippet: The proteins used in the study were prostate specific antigen (PSA) (P3338, Sigma-Aldrich), the spotted PSA antibody (ab403, Abcam), the Au NP PSA antibody (AF1344, R&D Systems), R-fetoprotein antigen (APF) (A32260H, Biodesign International), the spotted AFP antibody (10-A05, clone M19301, Fitzgerald Industries International, Inc.), the Au NP AFP antibody (70-XG05, Fitzgerald Industries International, Inc.), human chorionic gonadotropin (HCG) (A81355M, Biodesign International), the spotted HCG antibody (E20579, Biodesign International), and the Au NP-monoclonal HCG antibody (E20106, Biodesign International).

Techniques: Concentration Assay, Software

Journal: Advanced Science

Article Title: Conversion of Transplanted Mature Hepatocytes into Afp + Reprogrammed Cells for Liver Regeneration After Injury

doi: 10.1002/advs.202517126

Figure Lengend Snippet: Transplanted mature hepatocytes are reprogrammed at the early stage of liver regeneration. Additional details are provided in Table . (A) Schematic overview of the experimental strategy: serial transplantation of tdTomato + hepatocytes, followed by single‐cell sequencing of tdTomato + hepatocytes isolated from host livers at different timepoints post‐transplantation. (B) Uniform manifold approximation and projection (UMAP) of all 29,130 tdTomato + hepatocytes from eight samples, colored by cell clusters. Green and purple circles outline Group1 and Group2, respectively, as defined by hierarchical clustering ( left ). (C) Stream plot depicting the dynamic proportion distribution of Group1 and Group2 cells across eight samples. The width of each stream corresponds to the proportion of each group at different timepoints. (D) Volcano plot based on the percentage difference (x‐axis) and the log 2 (fold change) (log 2 FC, y‐axis) shows the upregulated genes in Group1 relative to Group2 cells. Genes highlighted in orange are important for identifying cellular states. (E) Donut chart showing the proportion of the four major categories of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enriched by the upregulated genes in Group1 relative to Group2 cells (adjusted p < 0.05). (F) Line charts display the average expression levels of three hepatic progenitor genes ( top ) and three cell cycle genes ( bottom ) over eight samples. Sample points represent the mean expression levels of all cells in a sample, error bars show the mean ± SEM, and segment bandwidths depict the confidence intervals. Asterisks mark significantly upregulated genes in each comparison (Tr_1 W vs. Tr_0 W and Tr_3 W vs. Tr_0 W; two‐sided Wilcoxon rank sum test, *** adjusted p < 0.001). (G) Scatter plot of the "Cell Cycle Score" (x‐axis) against the "Hepatocyte Maturation Score" (y‐axis). Green and purple dots correspond to Group1 and Group2 cells, respectively. The orange dotted line denotes the threshold set at the mean Cell Cycle Score plus three standard deviations, and cells above this threshold are classified as cycling cells. (H) Heatmap showing the average normalized enrichment score (NES) of pathways related to cellular stress response for each transplant timepoint relative to R0_0 W. Asterisks denote pathways that are significantly different from R0_0 W ( * adjusted p < 0.05, ** adjusted p < 0.01, *** adjusted p < 0.001). (I) Gene set enrichment analysis (GSEA) of reprogramming‐related genes (RRG) in Group1 vs. Group2 cells ( top ). Average NES of RRG for each transplant timepoint relative to R0_0 W ( bottom ). Asterisks mark a significant difference from R0_0 W ( * adjusted p < 0.05, *** adjusted p < 0.001). (J,K) Immunofluorescence (IF) staining of tdTomato and AFP (J) or Ki67 (K), respectively, in host livers at four timepoints post‐transplantation (1 W, 3 W, 6, and 12 W) ( left ). The ratios of tdTomato + AFP + (J) or tdTomato + Ki67 + (K) hepatocytes were quantified, respectively (shown as the bar chart to the right of each set of images). Scale bars: 200 µm. *** p < 0.001.

Article Snippet: Primary antibodies used in this study included: tdTomato (Arigo, ARG55724 , 1:400 dilution), AFP (Proteintech, 14550‐1‐AP, 1:200 dilution), E‐CAD (BD biosciences, 610181, 1:50 dilution), GS (Abcam, ab49873, 1:1000 dilution), Ki67(Abcam, 16667, 1:200 dilution), HNF4a (Abcam, 201460, 1:200 dilution), FAH (Cell Lab Tech, CLT‐602‐910, 1:3000 dilution), Images were acquired using a Leica microscope (m205), and were analyzed by the Image J.

Techniques: Transplantation Assay, Single Cell, Sequencing, Isolation, Expressing, Comparison, Immunofluorescence, Staining

Journal: Advanced Science

Article Title: Conversion of Transplanted Mature Hepatocytes into Afp + Reprogrammed Cells for Liver Regeneration After Injury

doi: 10.1002/advs.202517126

Figure Lengend Snippet: Metabolic remodeling and proliferation activation occur during the reprogramming of donor hepatocyte. Additional details are provided in Table . (A) Principal component analysis (PCA) plot based on maturation‐related genes encompassing 23 discrete timepoints across liver development and regeneration. The dataset includes fetal (E10.5‐E19.5) [ , ], postnatal (P0‐P60) , and transplanted (Adult/Tr_0W‐Tr_12 W) hepatocytes. Points are colored by their respective timepoint. Dotted lines in different colors outline the four major cell groups. (B‐C) PCA plot (B) and Diffusion map (C) each visualize the developmental progression of hepatocytes across 11 timepoints, including postnatal (P1‐P56) [ , ] and transplanted (Adult/Tr_0W‐Tr_12 W) stages. In both panels, cells are colored by timepoint. Inferred trajectories for maturation (black) and transplantation (red) are overlaid. (D) Donut chart showing the proportion of KEGG pathways enriched exclusively by the PC1‐related genes identified in Figure (adjusted p < 0.05). (E) Bar plot of KEGG pathways within the "Cell growth and death" category in Figure . The x‐axis represents the ‐log (adjusted p ) from the enrichment analysis. (F) Hierarchical clustering of hepatocytes across 12 timepoints segregated them into four groups based on global gene expression profiles. The 12 timepoints include the DDC_1 W sample , which was added as a distinct timepoint for comparison with the analyses in Figure . (G) Average module scores for gene sets characterizing hepatocyte states, shown for each group defined in Figure . Asterisks denote gene sets that are significantly different from the MH group (two‐sided Wilcoxon rank sum test, *** adjusted p < 0.001). (H) Dot plot showing the scaled expression (color intensity) and cellular percentage (dot size) of biliary genes that are expressed in ≥20% of cells in any group across the clusters defined in Figure . (I) Multiplex IF staining for tdTomato, AFP, and FAH ( left ) or CK19 ( right ) in host livers at 3 weeks post‐transplantation. Scale bars:100 µm. (J) UMAP plot ( bottom ) shows the module scores of the Afp + rHep‐related signature (ARS) for each cell. The ridge plot ( top ) shows the distribution of these scores for both cell groups. (K) Bar plot of the selected Gene Ontology (GO) and KEGG terms enriched for the ARS (adjusted p < 0.01). The x‐axis represents the ‐log (adjusted p ) of the enriched terms. (L) PCA plot of metabolic pathway enrichment scores in hepatocytes during early regeneration. Timepoints (n = 12) were selected based on clustering patterns in Figure , including PHx , CCl 4 , and APAP models (3 timepoints each), transplanted hepatocytes (2 timepoints), and LPLCs . Samples are colored by timepoint, revealing the exclusive localization of transplanted Afp + rHeps (Tr_1 W, Tr_3 W) in Quadrant I, indicative of their unique metabolic state. (M) Heatmap of NES for 82 KEGG metabolic pathways across the 12 regeneration samples. Color intensity indicates NES, revealing pathway‐level details underlying the PCA patterns in Figure . (N) Top 5 metabolic pathways with the highest specificity in Afp + rHeps. Bar plot shows the difference in mean enrichment scores between transplanted Afp + rHeps (Tr_1 W, Tr_3 W) and other regeneration hepatocytes (PHx, CCl 4 , APAP, LPLC) for pathways identified in Quadrant I of the pathway‐level PCA (Figure ). Positive values indicate Afp + rHeps‐specific enrichment. (O) Coordinated activation of energy metabolism in Afp + rHeps. Heatmap of Afp gene expression (mean) and NES for six core pathways shows specific upregulation of PPAR signaling (α/γ) , fatty acid oxidation (FAO) , TCA cycle, and oxidative phosphorylation (OXPHOS) in transplanted Afp + rHeps (Tr_1 W, Tr_3 W), indicating enhanced mitochondrial energy generation concurrent with high Afp expression.

Article Snippet: Primary antibodies used in this study included: tdTomato (Arigo, ARG55724 , 1:400 dilution), AFP (Proteintech, 14550‐1‐AP, 1:200 dilution), E‐CAD (BD biosciences, 610181, 1:50 dilution), GS (Abcam, ab49873, 1:1000 dilution), Ki67(Abcam, 16667, 1:200 dilution), HNF4a (Abcam, 201460, 1:200 dilution), FAH (Cell Lab Tech, CLT‐602‐910, 1:3000 dilution), Images were acquired using a Leica microscope (m205), and were analyzed by the Image J.

Techniques: Activation Assay, Diffusion-based Assay, Transplantation Assay, Gene Expression, Comparison, Expressing, Multiplex Assay, Staining, Phospho-proteomics

Journal: Advanced Science

Article Title: Conversion of Transplanted Mature Hepatocytes into Afp + Reprogrammed Cells for Liver Regeneration After Injury

doi: 10.1002/advs.202517126

Figure Lengend Snippet: AFP drives liver repair through PPAR‐centric transcriptional and metabolic networks in liver injury. Additional details are provided in Table . (A) Hierarchical clustering based on transcriptomic profiles distinguishes three distinct populations (IH, MH, RH) among postnatal (P1‐P56) and transplanted (Tr_1W‐Tr_3 W) hepatocytes. (B) Dot plot showing the scaled expression (color intensity) and cellular percentage (dot size) of Afp , H19 , and Cd63 across postnatal (P1‐P56) and transplanted hepatocytes (Tr_1W‐Tr_3 W). (C) Network diagrams of the top 10 genes (ranked by kME) within the hdWGCNA‐identified IH‐M2 module ( top , in IH group) and the RH‐M5 module ( bottom , in RH group). Node size is scaled by kME value, and edge thickness is weighted by connection strength, representing co‐expression relationships. The Afp gene is highlighted in blue. (D) Bar plot showing significantly enriched KEGG pathways shared between the Afp ‐associated gene modules IH‐M2 and RH‐M5 (adjusted p < 0.05). A dashed box denotes lipid metabolism‐related signaling. The x‐axis indicates the ‐log 10 (adjusted p ) of the enrichment analysis. (E) KEGG Pathways significantly enriched (adjusted p < 0.05) among the potential AFP‐interacting genes identified in both Ctl and AFP‐OE samples (listed in Table ). The x‐axis represents the ‐log 10 (adjusted p ‐value) for each pathway. (F) ( Top ) Representative IHC staining for FAH (a marker of donor repopulation) in liver sections of Fah −/− host mice with varying AFP backgrounds (Ctl, AFP‐OE, AFP‐KD) at 1, 2, and 3 weeks post‐transplantation with normal adult tdTomato + hepatocytes. Scale bars: 100 µm. ( Bottom )​ Quantification of the repopulation index, presented as the percentage of donor‐derived (tdTomato + ) area, in the three recipient groups over time. ** p < 0.01. (G) Kaplan‐Meier survival analysis of Fah −/− mice following hepatocyte transplantation, stratified by their systemic AFP background (Ctl, n = 4; AFP‐OE, n = 4; AFP‐KD, n = 8). * p < 0.05 (AFP‐KD vs. Ctl/AFP‐OE). (H) ( Left ) Representative IHC staining of AFP in liver sections. Mice were treated with CCl 4 alone or CCl 4 combined with rAFP injection, and tissues were harvested at 2, 3, 4, and 7 days post‑administration. Scale bars: 100 µm. ( Right ) Quantification of AFP + cells, presented as the percentage of AFP + cells per field, in the two treatment groups across the indicated timepoints. ns: not significant, * p < 0.05, *** p < 0.001. (I) Representative H&E staining and quantification of necrotic area in liver sections from mice at 2 days postadministration. Mice were treated with CCl 4 alone or CCl 4 combined with rAFP injection. Scale bars: 100 µm. *** p < 0.001. (J) Representative H&E‐stained liver sections from mice 48 h after intraperitoneal injection of 300 mg/kg APAP. Mice were concurrently treated with either vehicle (Control), low‐dose rAFP, or high‐dose rAFP. Necrosis is markedly reduced in rAFP‐treated livers ( *** p < 0.001). Scale bars: 100 µm. (K) Kaplan‐Meier survival curves following a single intraperitoneal injection of a lethal dose of CCl 4 (3 mL/kg). Mice were treated with either CCl 4 alone (Control) or CCl 4 followed by rAFP treatment. * p < 0.05. (L) Kaplan‐Meier survival curves of mice following a lethal dose of APAP (800 mg/kg) with or without rAFP treatment (based on the circulating blood volume of mice). * p < 0.05, *** p < 0.001. (M) Quantitative RT‐PCR (qRT‐PCR) analysis of gene expression in primary mouse hepatocytes treated for 24 h with: control, rAFP alone, rAFP combined with the PPARγ agonist Rosiglitazone (10 µ m ), or rAFP combined with the PPARγ antagonist GW9662 (10 µ m ). mRNA levels were assessed for the Afp gene and four canonical downstream genes of the PPARγ signaling pathway ( Cd36 , Fabp4 , Fasn , Scd1 ). * p < 0.05, ** p < 0.01, *** p < 0.001. (N) ( Left ) Representative FAH IHC staining of liver sections from Fah −/− mice at 3 weeks post‐transplantation. Mice were treated as indicated: Basal, rAFP, or rAFP+GW9662. Scale bars: 200 µm. ( Right ) Quantification of the repopulation index, presented as the percentage of donor‐derived (FAH + ) area, in the three treatment groups. *** p < 0.001.

Article Snippet: Primary antibodies used in this study included: tdTomato (Arigo, ARG55724 , 1:400 dilution), AFP (Proteintech, 14550‐1‐AP, 1:200 dilution), E‐CAD (BD biosciences, 610181, 1:50 dilution), GS (Abcam, ab49873, 1:1000 dilution), Ki67(Abcam, 16667, 1:200 dilution), HNF4a (Abcam, 201460, 1:200 dilution), FAH (Cell Lab Tech, CLT‐602‐910, 1:3000 dilution), Images were acquired using a Leica microscope (m205), and were analyzed by the Image J.

Techniques: Expressing, Immunohistochemistry, Marker, Transplantation Assay, Derivative Assay, Injection, Staining, Control, Quantitative RT-PCR, Gene Expression

Journal: Advanced Science

Article Title: Conversion of Transplanted Mature Hepatocytes into Afp + Reprogrammed Cells for Liver Regeneration After Injury

doi: 10.1002/advs.202517126

Figure Lengend Snippet: Zonation reconstruction of the host liver by the transplanted mature hepatocytes. Additional details are provided in Table . (A) UMAP visualization of transplanted hepatocytes from two rounds, collected at 1, 3, 6, and 12 weeks post‐transplantation, colored by model‐predicted cell zonation (Zone 1–3). (B) Stacked bar plot showing the distribution of predicted hepatocyte zonation (Zone1‐3) across eight samples. (C) IF staining for tdTomato, GS, and E‐CAD in livers at 1, 3, 6, and 12 weeks post‐transplantation ( top ). Zoom in on the selected field of view ( bottom ). Scale bars: 100 µm. (D) Lollipop plot of top 5 KEGG pathways most closely matched to hepatocyte zonation dynamics (across eight samples) as quantified by Dynamic Time Warping (DTW) distance. Smaller DTW values indicate stronger alignment between pathway activity and zonation pattern changes. (E) Line plots illustrate the temporal dynamics of JAK‐STAT ( top ) and PPAR ( bottom ) pathway activities (GSVA scores) across hepatocyte zonation (Zone1‐3). Data points represent the mean scores of zonated hepatocytes within each group (stratified by zone and timepoint), and error bars denote the mean ± SEM. (F) Line plot showing the temporal dynamics of TGFb, MAPK, and PI3K pathway activities (inferred by PROGENy ) across eight samples. Data points represent the mean activity of hepatocytes in each sample, and error bars denote the mean ± SEM. Asterisks denote significantly upregulated pathways in each comparison (R1_1 W/3 W vs. R0_0 W and R2_3 W vs. R1_12 W; two‐sided Wilcoxon rank sum test, *** adjusted p < 0.001). (G) Heatmap showing the average NES of pathways related to cell junction and cell polarity for hepatocytes at each transplant timepoint relative to Adult hepatocytes. Asterisks mark significant enrichment ( * adjusted p < 0.05, ** adjusted p < 0.01, *** adjusted p < 0.001). (H) Dot plot showing the scaled expression (color intensity) and cellular percentage (dot size) of epithelial mesenchymal transition (EMT)‐related genes in Afp + rHep and MH. (I) IF staining for tdTomato, AFP, and CK18 ( left ) or FN1 ( right ) in livers at 3 weeks post‐transplantation. Scale bars: 100 µm. (J) GSEA of epithelial mesenchymal plasticity (EMP) signature in Afp + rHep vs. MH.

Article Snippet: Primary antibodies used in this study included: tdTomato (Arigo, ARG55724 , 1:400 dilution), AFP (Proteintech, 14550‐1‐AP, 1:200 dilution), E‐CAD (BD biosciences, 610181, 1:50 dilution), GS (Abcam, ab49873, 1:1000 dilution), Ki67(Abcam, 16667, 1:200 dilution), HNF4a (Abcam, 201460, 1:200 dilution), FAH (Cell Lab Tech, CLT‐602‐910, 1:3000 dilution), Images were acquired using a Leica microscope (m205), and were analyzed by the Image J.

Techniques: Transplantation Assay, Staining, Activity Assay, Comparison, Expressing