Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: Intestinal DCs induce Foxp3 expression of effector T cells with higher efficiency than splenic DC. (A) 10 5 Foxp3 − cells were cultured with 2 × 10 4 splenic or LpDCs for 5 d with α-CD3 mAb alone or combined with TGF-β. Plots are gated on CD4 + cells, and the percentages of Foxp3 + cells are shown. Phenotypic analysis of converted T reg cells is shown on the right. 5-d cultured cells from A were stained for CD4 and CD103. Histogram shows the CD103 expression of CD4 + Foxp3 + cells cultured with SpDCs (thick line) or LpDCs (thin line). (B) 10 5 Foxp3 − CD4 + T cells cultured with DCs and α-CD3 mAb and TGF-β as in (A) for 5 d, with the ratio of T/DC as 5/1, 10/1, 20/1 or 40/1. Data are the mean ± SD of triplicate wells. (C) Foxp3 − cells were cultured as in A with CD103 + LpDC or CD103 − LpDC. Plots are gated on CD4 + cells and Foxp3 versus α 4 β 7 stainings are shown. The numbers indicate percentage of events in each quadrant. Data are from one of at least three independent experiments.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Expressing, Cell Culture, Staining

Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: Maintenance of Foxp3 contributes to higher frequency of Foxp3 + CD4 + T cells in the presence of LpDC. (A) eGFP − CD4 + T cells were cocultured with purified DC at a 5:1 ratio with α-CD3 and TGF-β. At indicated time points, cells were harvested and stained for CD4, Class II, and 7-AAD. Foxp3 expression was determined based on eGFP fluorescence. The percentage of viable cells is depicted by open bars for LpDC and filled bars for SpDC. Data are from one of two independent experiments. (B) CFSE-labeled naive CD4 + T cells were cocultured with purified DCs as described in A. On day 5, cells were harvested and stained for CD4 and intracellular Foxp3. Illustrated are dot plots of Foxp3 versus CFSE. The percentage of Foxp3 + cells and the MFI was defined as the bordered population. (C) In cultures containing LpDC (○) or SpDC (•) the proportion of Foxp3 + cells among CD4 T cells is plotted as a function of the number of cell divisions. Data are representative of three independent experiments.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Purification, Staining, Expressing, Fluorescence, Labeling

Journal: The Journal of Experimental Medicine

Article Title: Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid

doi: 10.1084/jem.20070602

Figure Lengend Snippet: RA production by LpDC is requisite for optimal Foxp3 + T reg cell conversion. Foxp3 − CD4 T cells were cocultured with DC at a 10:1 ratio as described in Cells were stained for α 4 β 7 and assessed for eGFP fluorescence (Foxp3) by flow cytometry. Results are representative of three independent experiments. (B) The dose responsiveness of all-trans RA on eGFP (Foxp3) expression by CD4 T cells cocultured with SpDC was determined by flow cytometry. Error bars represent the SDs of the means of three individual samples from one experiment. Statistical significance was determined using the Student's t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Results are representative of three independent experiments. (C) Foxp3 − CD4 + T cells were cocultured with CD103 + or CD103 − DC at a 10:1 ratio as described in . Cells were stained for α 4 β 7 and the effect of 100 nM RA, or the RA receptor inhibitors LE540 and LE135 on (eGFP) Foxp3 expression was determined by flow cytometry. Results are representative of three independent experiments. (D) Foxp3 − CD4 T cells were cocultured with CD103 + DC in the presence of α-CD3 or α-CD3 in combination with TGF-β neutralizing antibody (isotype IgG1) or LE540 and LE135. eGFP (Foxp3) expression was determined by flow cytometry. NS. Error bars represent the SD from the means of three independent experiments. ***, P < 0.001.

Article Snippet: In some experiments, the following were included in the coculture conditions: (a) all-trans RA (Sigma-Aldrich) at varying concentrations; (b) RA receptor β inhibitors LE540 (Wako Chemicals USA) and LE135 (Tocris Bioscience), each at 1 μM; and (c) α-TGF-β mAb (1D11.16.8) or isotype control mAb MOPC (31C; both from American Type Culture Collection) at 10 μg/ml.

Techniques: Staining, Fluorescence, Flow Cytometry, Expressing

Journal: The Journal of Immunology Author Choice

Article Title: Generation of an Inhibitory NK Cell Subset by TGF-β1/IL-15 Polarization

doi: 10.4049/jimmunol.2300834

Figure Lengend Snippet: TGF-β1 and IL-15 induce CD103 and CD49a expression on NK cells. PBMCs were stimulated with TGF-β1 and IL-15 or with IL-15 alone for 7 d in vitro and stained for CD103. (A) Contour plot showing percentage of CD56+Lin− (CD3−CD14−CD19−CD20−CD34−CD123−CD303−FCεRIα−TCRαβ−TCRγδ−) NK cells expressing CD103. U/S PB NK cells are also shown as a control condition. A total of five experiments across six PB donors were conducted. (B and C) Percentage of CD56+Lin− NK cells expressing CD103 after NK cells were stimulated with different cytokine combinations, including (B) IL-2, IL-7, IL-12, IL-18, and IL-21 in combination with TGF-β1 and (C) IL-15 in combination with different cytokines. U/S represents U/S CD56+Lin− NK cells. Data are representative of two independent experiments with two different donors. (D) Representative contour plots from PB NK cells stimulated with TGF-β1 and IL-15, IL-15 only, or U/S showing expression of CD103 versus CD49a or CD103 versus CD69. Figures are representative of four separate experiments conducted across five PB donors. Two-way ANOVA (main effect of TGF-β1, p < 0.0001) (C) and Welch’s t test (B) were used for statistical analysis. ***p < 0.001; ****p < 0.0001. Variances are all displayed as SEM unless otherwise specified.

Article Snippet: In some experiments, cells were treated with or without anti-TGF-β neutralizing Ab (10 μg/ml, catalog no. BP0057, 1D11.16.8, Bio X Cell).

Techniques: Expressing, In Vitro, Staining, Control

Journal: The Journal of Immunology Author Choice

Article Title: Generation of an Inhibitory NK Cell Subset by TGF-β1/IL-15 Polarization

doi: 10.4049/jimmunol.2300834

Figure Lengend Snippet: Ascites-derived TGF-β induces expression of CD103 and CD49a on PB NK cells. (A) To determine whether the TME induces regulatory features in NK cells, PB NK cells from healthy donors were cultured with 50% supernatant of ascites from patients with EOC and IL-15 for 7 d. (B) Contour plots of a sample where PB CD56+Lin− NK cells were stimulated with 50% ascites supernatant and IL-15 versus U/S. Expression of CD103 with either CD49a (top panels) or CD69 (bottom panels) are shown. Patient-matched ascites CD56+ NK cells are also shown (right panels). Coexpression analysis was done across two ascites samples (i.e., ASC641, ASC888). (C) Dot plots showing expression of CD103, CD49a, or CD69 in U/S PB NK cells compared with paired PB NK cells stimulated with 50% ascites supernatant and IL-15 from multiple patients (n = 4–5; i.e., ASC591, ASC635, ASC640, ASC641, ASC888). Paired Wilcoxon test or Student t test was performed. (D) Ascites-stimulated PB CD56+ ILCs treated with or without anti-TGF-β neutralizing Abs for 7 d. Supernatant from six ascites samples was used (i.e., ASC736, ASC743, ASC748, ASC769, ASC775, ASC777). The percentage of cells positive for CD103 is shown. Two-way ANOVA (F[1,12] = 76.80) and multiple comparisons with Dunn-Šidák correction was performed. Data included two or three technical replicates. Data with supernatant ASC777 was repeated in a separate experiment. (E) Summary plot and pairwise comparisons of (D) with lines representing matched ascites supernatants. Variance displayed as min/max, and unpaired Student t test was used for statistical significance. TME, tumor microenvironment. *p < 0.05; ***p < 0.001; ****p < 0.0001.

Article Snippet: In some experiments, cells were treated with or without anti-TGF-β neutralizing Ab (10 μg/ml, catalog no. BP0057, 1D11.16.8, Bio X Cell).

Techniques: Derivative Assay, Expressing, Cell Culture

Journal: The Journal of Immunology Author Choice

Article Title: Generation of an Inhibitory NK Cell Subset by TGF-β1/IL-15 Polarization

doi: 10.4049/jimmunol.2300834

Figure Lengend Snippet: TGF-β1/IL-15–induced NK-like cells express markers similar to intratumoral CD103+CD56+ ILCs. (A) Time kinetics of surface markers and transcription factors expressed by PB CD56+Lin− ILCs that were stimulated with IL-15 and TGF-β1 for 1, 3, and 7 d. Circle plots show proportion of PB CD56+Lin− NK cells coexpressing select markers throughout time averaged across two technical replicates. Numbers in square brackets represent the number of markers coexpressed. Findings are representative of two separate experiments across three different PB donors. (B) Circle plots comparing coexpression of markers by CD56+ NK/ILCs from PB from healthy donor, ascites from EOC patients, or EOC tumors. Each circle plot represents an individual sample. Representative contour plots of CD56+Lin− NK/ILCs from healthy PB donors, tumor, or PB NK cells cultured with IL-15 and TGF-β1 for 7 d (C and D). Expression of CD103 in combination with CD49a or CD69 (C), or CD101 or GITR (D), are shown. Findings for IL-15/TGF-β1 stimulation are representative of three separate experiments.

Article Snippet: In some experiments, cells were treated with or without anti-TGF-β neutralizing Ab (10 μg/ml, catalog no. BP0057, 1D11.16.8, Bio X Cell).

Techniques: Cell Culture, Expressing

Journal: The Journal of Immunology Author Choice

Article Title: Generation of an Inhibitory NK Cell Subset by TGF-β1/IL-15 Polarization

doi: 10.4049/jimmunol.2300834

Figure Lengend Snippet: TGF-β1/IL-15–induced CD103+ NK-like cells suppressed autologous CD4+ T cells in vitro. To determine whether TGF-β1/IL-15–induced CD103+ NK-like cells are functionally inhibitory, TGF-β1/IL-15–induced CD103+ NK-like cells were cocultured with autologous CD4+ T cells. (A) Schematic diagram of suppression assay using TGF-β1/IL-15–induced CD103+ NK-like cells. PB CD56+ NK cells were stimulated with IL-15 and TGF-β1 for 7 d. TGF-β1/IL-15–induced CD103+CD56+Lin− (CD3−CD14−CD19−) NK-like cells were FACS sorted and cocultured in 1:1 ratio with autologous CD4+ T cells stimulated with Dynabeads (coated with αCD3/αCD28) for 4 d. These assays have two or three technical replicates, and data are representative of three independent experiments. (B) Absolute numbers of live CD4+ T cells that remained after coculture with either TGF-β1/IL-15–induced CD103+ NK-like cells, U/S NK cells, or T cell only in vitro with different cytokine conditions (i.e., IL-2, IL-15, or no cytokines). Two-way ANOVA with Dunn-Šidák multiple comparisons test was performed for CD4+ T cells (main effect of cells cocultured; F[2,17] = 85.79, p < 0.0001). (C) Percentage suppression was calculated using absolute CD4+ T cell numbers after exposure to either U/S NK cells or TGF-β1/IL-15–induced CD103+ NK-like cells, normalized to T cell only conditions. Suppression assay shown has a decreasing ratio of suppressors to responder cells from 1:1 to 1:8. (D) Proliferation of live CD4+ T cells with either TGF-β1/IL-15–induced CD103+ NK-like cells, U/S NK cells, or T cell only using cell trace proliferation dye. Expression of activation marker CD25 is shown from CD4+ T cells at a 1:1 ratio (E) or a downward titration (F). (G) Percentage of dead CD4+ T cells measured using fixable viability dye. (H) Supernatant from cocultures at day 4 were collected for CBA assay and concentration of perforin is shown. Data are representative of two separate experiments. Two-way ANOVA with Dunn-Šidák multiple comparisons test was performed for (C) (main effect of cells cocultured, F[1,11] = 107, p < 0.0001), (F) (main effect of cells cocultured, F[1,8] = 128.4, p < 0.0001), and (H) (main effector of cells cocultured, F[1,8] = 184.5, p < 0.0001). One-way ANOVA with Dunnett’s multiple comparison test was performed for (E) (F[2,6] = 8.403, p = 0.0182) and (G) (F[2,5] = 120.7, p < 0.0001). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; variance displayed as SEM. CBA, cytometric bead array.

Article Snippet: In some experiments, cells were treated with or without anti-TGF-β neutralizing Ab (10 μg/ml, catalog no. BP0057, 1D11.16.8, Bio X Cell).

Techniques: In Vitro, Suppression Assay, Expressing, Activation Assay, Marker, Titration, Concentration Assay, Comparison

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Detection and Significance of Cytotoxic Cell Subsets in Biopsies of HCV-Infected Human Livers

doi: 10.1007/s00005-013-0258-6

Figure Lengend Snippet: Primary antibodies used

Article Snippet: NKG2D/CD314 , Mouse , Monoclonal , 1D11 , Novus Biologicals , NB100-65956.

Techniques:

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Slit3 repression correlated with HCC tumor growth. a Tumor size of HCC patients cohort 1 ( n = 40) with high or low Slit3 expression. b Relative Slit3 expression inversely correlated with tumor size of HCC patients cohort 1 ( n = 40). c Tumor size of HCC patients cohort 2 ( n = 25) with high or low Slit3 expression. d Relative Slit3 expression inversely correlated with tumor size of HCC patients cohort 2 ( n = 25)

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Slit3 negatively regulated HCC cell growth. a Expression of Slit3 in HCC cell-lines. b and c Stable Slit3 repressed clones (shSlit3) of LM3 and PLC cells displayed induced cell proliferation when compared with the control cells (shCTL) after 72 h. d Stable Slit3 overexpressed clones (Slit3) of Hep3B cells displayed impaired cell proliferation when compared with the control cells (pcDNA) after 72 h. The protein expression level was quantified by ImageJ software and normalized to the expression of actin. The experiment was repeated for three times and one representative blot was shown

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing, Clone Assay, Control, Software

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Molecular mechanism associated with stable Slit3 repression. Protein expression of certain proteins associated with cell cycle progression (cyclin D3), apoptosis (survivin) and GSK3β/β-catenin pathway (β-catenin and phosopho-GSK3β (ser9) in LM3 and PLC Slit3 shRNA transfectants (shSlit3) and shRNA control (shCTL). The protein expression level was quantified by ImageJ software and normalized to the expression of actin. The experiment was repeated for three times and one representative blot was shown

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing, shRNA, Control, Software

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Slit3 negatively regulated HCC cell growth in vivo. a Stable Slit3 repressed clones (LM3-shSlit3) displayed induced tumor growth when compared with the control cells (LM-shCTL) 6 weeks post-injection of cells subcutaneously into flank region of nude mice ( n = 5). b Stable Slit3 overexpressed clones (Hep3B-Slit3) displayed impaired tumor growth when compared with the control cells (Hep3B-pcDNA) 6 weeks post-injection of cells subcutaneously into flank region of nude mice ( n = 5). c Immunohistochemical staining of Slit3, CD31, p27, β-catenin and phosopho-GSK3β (ser9) in tumor formed by LM3-shCTL and LM3-shSlit3 stable cells. d Immunohistochemical staining of Slit3, CD31, p27, β-catenin and phosopho-GSK3β (ser9) in tumor formed by Hep3B-pcDNA and Hep3B-Slit3 stable cells. Representative results from two mice of each group were shown

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: In Vivo, Clone Assay, Control, Injection, Immunohistochemical staining, Staining

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Stable repression of Slit3 expression enhanced chemoresistance in HCC cells. (Left panel) LM3-shSlit3 and shCTL cells and (right panel) PLC-shSlit3 and shCTL cells were treated with ( a ) 10 μM sorafenib, ( b ) 10 μM oxaliplatin and ( c ) 100 μM 5-FU for 72 h. The number of cells was determined by MTT assay and expressed as percentage of that under vehicle control treatment. When compared with shCTL cells, shSlit3 cells showed significantly stronger resistance to the treatment of all chemotherapeutic drugs. Each experiment was performed in triplicate and data was obtained from three independent experiments

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing, MTT Assay, Control

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Molecular mechanism associated with Slit3 repression-induced chemoresistance. Protein expression of β-catenin, cyclin D3 and survivin in LM3 and PLC Slit3 shRNA transfectants (shSlit3) and shRNA control (shCTL) following 72-h vehicle control (C), 10 μM sorafenib (SOR), 10 μM oxaliplatin (OXA) and 100 μM 5-FU (5FU) treatment. The protein expression level was quantified by ImageJ software and normalized to the expression of actin. The experiment was repeated for three times and one representative blot was shown

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing, shRNA, Control, Software

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Down-regulation of β-catenin diminished the effects of Slit3-repression. Stable PLC Slit3-repressed cells (PLC-shSlit3) and control clone (PLC-shCTL) were transiently transfected with siRNA control (siCTL) or β-catenin siRNA (si-β-catenin) to investigate the effect of β-catenin down-regulation on Slit3-regulated effects. a PLC-shSlit3 siCTL cells showed significantly higher proliferation rate than PLC-shCTL siCTL cells whereas such induction was significantly impaired in PLC-shSlit3 si-β-catenin cells. b to d The induced chemoresistance in PLC-shSlit3 siCTL cells when compared with PLC-shCTL siCTL cells upon treatment of sorafenib, oxaliplatin and 5-FU were impaired in PLC-shSlit3 si-β-catenin cells. e The inductions in protein expression of phospho-GSK3β, cyclin D3 and survivin in PLC-shSlit3 siCTL cells when compared with PLC-shCTL siCTL were all reduced in PLC-shSlit3 si-β-catenin cells

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Control, Transfection, Expressing

Journal: BMC Cancer

Article Title: Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway

doi: 10.1186/s12885-018-4326-5

Figure Lengend Snippet: Clinicopathological correlation of Slit3 expression in HCC patients

Article Snippet: Antibody against Slit3 was purchased from Novus Biologicals (Littleton, CO).

Techniques: Expressing, Over Expression, Infection

Journal: Advanced Science

Article Title: Cilia‐Mediated Insulin/Akt and ST2/JNK Signaling Pathways Regulate the Recovery of Muscle Injury

doi: 10.1002/advs.202202632

Figure Lengend Snippet: IL13 expression in FAPs is upregulated in Tchp −/− mice. A) qRT‐PCR analysis of RNA isolated from whole TA muscle of WT and Tchp −/− mice ( n = 3 or 4) at 3 dpi. B) Immunofluorescence for IL13 (red), hematopoietic cells (CD45, green), and nuclei (Hoechst33342, blue) in TA muscle cross‐sections of WT and Tchp −/− mice at 3 dpi. Lower magnification images are shown in Figure S6A, Supporting Information. Scale bars, 10 µm. C) Quantification of the IL4‐, IL13‐, or IL33‐stained area per 40× view in (B) and Figure S6A,B (Supporting Information) ( n = 6 or 7 fields each). D) Quantification of the IL13‐stained area per 40× view excluding the CD45‐stained area in (B) ( n = 7 fields each). E) Immunofluorescence for IL4 (red), IL13 (red), and IL33 (red), PDGFR α + (green), and nuclei (Hoechst33342, blue) in PDGFR α + FAPs isolated from hindlimbs of WT and Tchp −/‐ mice. Scale bars: 20 µm. All data are the mean ± S.D from 3 to 5 mice. * p < 0.05, ** p < 0.01, N.S., not significant; two‐tailed unpaired Student's t ‐tests.

Article Snippet: Staining was performed using the following antibodies: acetylated tubulin (6‐11B‐1, Sigma–Aldrich; 1:200), Arl13b (17711‐1‐AP, Proteintech; 1:200), PTRF (18892‐1‐AP, Proteintech; 1:100), Caveolin‐1 (#3238, Cell Signaling Technology; 1:300), Collagen I (ab21286, Abcam; 1:100), Perilipin (D1D8, Cell Signaling Technology; 1:200), CD45 and CD45‐biotin (30‐F11, Biolegend; 1:100), CD68 (FA‐11, Bio‐Rad; 1:100), CD206 (MR5D3, Bio‐Rad; 1:100), Desmin (D93F5, Cell Signaling Technology; 1:200), Flotillin‐2 (B‐6, Santa Cruz Biotechnology; 1:20), FOXP3 (FJK‐16s, Invitrogen; 1:100), GM3 (GMR6, Tokyo Chemical Industry; 1:500), IL4 (BVD4‐1D11, Novus Biologicals; 1:100), IL13 (AF‐413, R&D Systems; 1:100), IL33 (AF3626, R&D Systems; 1:100), PAX7 (MAB1675, R&D Systems; 1:100), PDGFR α (AF1062, R&D Systems; 1:200; APA5, Biolegend; 1:100), p‐Histone H3 (Ser 10) (sc‐8656‐R, Santa Cruz Biotechnology; 1:100), Sca1 (E13‐161.7, Biolegend, 1:100), Vimentin (#280618, R&D Systems; 1:200), Ki67 (SolA15, Invitrogen; 1:200), and Laminin (L9393, Sigma Aldrich; 1:200).

Techniques: Expressing, Quantitative RT-PCR, Isolation, Immunofluorescence, Staining, Two Tailed Test