Journal: Cell stem cell

Article Title: A scalable organoid model of human autosomal dominant polycystic kidney disease for disease mechanism and drug discovery

doi: 10.1016/j.stem.2022.06.005

Figure Lengend Snippet: Key Resources Table

Article Snippet: The following probes from ThermoFisher were used for transcriptional analyses: WT1 (Hs01103751_m1) , MAFB (Hs00534343_s1) , PAX2 (Hs01057416_m1) , HNF4A (Hs00230853_m1) , GATA3 (Hs00231122_m1) , SLC3A1 (Hs00942976_m1) , SLC12A1 (Hs00165731_m1) and SLC12A3 (Hs01027568_m1). . e) Western Blot To prepare protein lysate samples, organoids and control samples were suspended and homogenized in lysis buffer (RIPA buffer (Pierce, 89901) supplemented with 1 mM benzamidine hydrochloride (TCI America, TCB0013), 1X protease inhibitor cocktail (Cell signaling, 5871), 100 μM PMSF (Sigma-Aldrich, 11359061001), and protease inhibitor cocktail tablets (one tablet/10ml of buffer) (Sigma, 11836170001)), and left on ice for 30mins.

Techniques: Plasmid Preparation, Recombinant, TUNEL Assay, Mutagenesis, Software

Journal: Cell stem cell

Article Title: A scalable organoid model of human autosomal dominant polycystic kidney disease for disease mechanism and drug discovery

doi: 10.1016/j.stem.2022.06.005

Figure Lengend Snippet: Key Resources Table

Article Snippet: The following probes from ThermoFisher were used for transcriptional analyses: WT1 (Hs01103751_m1) , MAFB (Hs00534343_s1) , PAX2 (Hs01057416_m1) , HNF4A (Hs00230853_m1) , GATA3 (Hs00231122_m1) , SLC3A1 (Hs00942976_m1) , SLC12A1 (Hs00165731_m1) and SLC12A3 (Hs01027568_m1). . e) Western Blot To prepare protein lysate samples, organoids and control samples were suspended and homogenized in lysis buffer (RIPA buffer (Pierce, 89901) supplemented with 1 mM benzamidine hydrochloride (TCI America, TCB0013), 1X protease inhibitor cocktail (Cell signaling, 5871), 100 μM PMSF (Sigma-Aldrich, 11359061001), and protease inhibitor cocktail tablets (one tablet/10ml of buffer) (Sigma, 11836170001)), and left on ice for 30mins.

Techniques: Plasmid Preparation, Recombinant, TUNEL Assay, Mutagenesis, Software

Journal: Neoplasia (New York, N.Y.)

Article Title: Nephron Progenitor But Not Stromal Progenitor Cells Give Rise to Wilms Tumors in Mouse Models with β-Catenin Activation or Wt1 Ablation and Igf2 Upregulation 1

doi: 10.1016/j.neo.2015.12.001

Figure Lengend Snippet: Summary of Mouse Mutant Cohorts and TM Injection Conditions

Article Snippet: Antibodies used were: WT1 (sc-192, Santa Cruz Biotechnology), Ki67 (ab15580, Abcam), pHH3 (06-570, Upstate Biotechnology), β-catenin (610154, BD Biosciences), Dlk1 (sc-8624, Santa Cruz Biotechnology), Cited1 (9219, Fisher Scientific), Six2 (11562-1-AP, Proteintech), Pax2 (PRB-276P, Covance), NCAM (C9672, Sigma), E-cadherin (3195, Cell Signaling Technology), K-cadherin (ab79005, Abcam), Vimentin (V2258, Sigma), Collagen IV (AB756P, Chemicon), Cyclin D1 (sc-753, Santa Cruz Biotechnology), and C-myc (9E10, Santa Cruz Biotechnology).

Techniques: Mutagenesis, Injection

Journal: Neoplasia (New York, N.Y.)

Article Title: Nephron Progenitor But Not Stromal Progenitor Cells Give Rise to Wilms Tumors in Mouse Models with β-Catenin Activation or Wt1 Ablation and Igf2 Upregulation 1

doi: 10.1016/j.neo.2015.12.001

Figure Lengend Snippet: Development of WT from nephron progenitors. (A) Kaplan-Meier tumor-free survival analysis showing incidence of WT in Six2 GCE , Cited1 Cre , and Foxd1 GCE mutant mice with Wt1 ablation/haploinsufficiency and β-catenin stabilization ( S/C/F-Wt1 −/fl -β-cat S and S/C/F-Wt1 +/fl -β-cat S ) and Wt1 ablation and Igf2 upregulation ( S/C/F-Wt1 −/fl -Igf2 ) in comparison to control mice. (B) PCR analysis of DNA from mutant mice showing Ctnnb1 wild-type or exon3 floxed allele and exon3 deleted allele ( Ctnnb1 + or Ctnnb1 ex3(fl) and Ctnnb1 ex3Δ ) and Wt1 floxed or null alleles (Wt1 fl or Wt1 Δ ).

Article Snippet: Antibodies used were: WT1 (sc-192, Santa Cruz Biotechnology), Ki67 (ab15580, Abcam), pHH3 (06-570, Upstate Biotechnology), β-catenin (610154, BD Biosciences), Dlk1 (sc-8624, Santa Cruz Biotechnology), Cited1 (9219, Fisher Scientific), Six2 (11562-1-AP, Proteintech), Pax2 (PRB-276P, Covance), NCAM (C9672, Sigma), E-cadherin (3195, Cell Signaling Technology), K-cadherin (ab79005, Abcam), Vimentin (V2258, Sigma), Collagen IV (AB756P, Chemicon), Cyclin D1 (sc-753, Santa Cruz Biotechnology), and C-myc (9E10, Santa Cruz Biotechnology).

Techniques: Mutagenesis, Comparison, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Nephron Progenitor But Not Stromal Progenitor Cells Give Rise to Wilms Tumors in Mouse Models with β-Catenin Activation or Wt1 Ablation and Igf2 Upregulation 1

doi: 10.1016/j.neo.2015.12.001

Figure Lengend Snippet: Histological analysis of WTs from Six2 GCE and Cited1 Cre mice. (A) Gross appearance of tumors from Six2 GCE mice (a) and Cited1 Cre mice (b and c) of the indicated genotypes. (B) H&E staining of kidney tumor sections of S-Wt1-β-cat S , C-Wt1-β-cat S , and C-Wt1-Igf2 mice. Scale bar: 100 μm.

Article Snippet: Antibodies used were: WT1 (sc-192, Santa Cruz Biotechnology), Ki67 (ab15580, Abcam), pHH3 (06-570, Upstate Biotechnology), β-catenin (610154, BD Biosciences), Dlk1 (sc-8624, Santa Cruz Biotechnology), Cited1 (9219, Fisher Scientific), Six2 (11562-1-AP, Proteintech), Pax2 (PRB-276P, Covance), NCAM (C9672, Sigma), E-cadherin (3195, Cell Signaling Technology), K-cadherin (ab79005, Abcam), Vimentin (V2258, Sigma), Collagen IV (AB756P, Chemicon), Cyclin D1 (sc-753, Santa Cruz Biotechnology), and C-myc (9E10, Santa Cruz Biotechnology).

Techniques: Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Nephron Progenitor But Not Stromal Progenitor Cells Give Rise to Wilms Tumors in Mouse Models with β-Catenin Activation or Wt1 Ablation and Igf2 Upregulation 1

doi: 10.1016/j.neo.2015.12.001

Figure Lengend Snippet: Activity of Wnt/β-catenin signaling in mouse WTs. (A) IHC staining of β-catenin and C-myc for sections from control kidneys and tumors from S-Wt1-β-cat S , C-Wt1-β-cat S , and C-Wt1-Igf2 mutants. Scale bar: 100 μm. (B) qPCR analysis of Wnt/β-catenin canonical effectors Axin2 , CyclinD1 , and C-myc and signaling inhibitors Wif1 and Dkk2 in littermate kidneys and tumors from S-Wt1-β-cat S , C-Wt1-β-cat S , and C-Wt1-Igf2 mutants. The x -axis labels indicate wild-type (Wt1 +/fl ) in littermate kidneys and the presence of progenitor-specific Cre alleles ( Six2 GCE or Cited1 Cre ) or Ubiquitous-Cre (U) and the presence of genetic alterations of β-cat S , Wt1 ablation ( Wt1 −/∆ ) or Igf2 upregulation ( H19 +/− m ) in mouse tumors.

Article Snippet: Antibodies used were: WT1 (sc-192, Santa Cruz Biotechnology), Ki67 (ab15580, Abcam), pHH3 (06-570, Upstate Biotechnology), β-catenin (610154, BD Biosciences), Dlk1 (sc-8624, Santa Cruz Biotechnology), Cited1 (9219, Fisher Scientific), Six2 (11562-1-AP, Proteintech), Pax2 (PRB-276P, Covance), NCAM (C9672, Sigma), E-cadherin (3195, Cell Signaling Technology), K-cadherin (ab79005, Abcam), Vimentin (V2258, Sigma), Collagen IV (AB756P, Chemicon), Cyclin D1 (sc-753, Santa Cruz Biotechnology), and C-myc (9E10, Santa Cruz Biotechnology).

Techniques: Activity Assay, Immunohistochemistry, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Nephron Progenitor But Not Stromal Progenitor Cells Give Rise to Wilms Tumors in Mouse Models with β-Catenin Activation or Wt1 Ablation and Igf2 Upregulation 1

doi: 10.1016/j.neo.2015.12.001

Figure Lengend Snippet: Expression of differentiation markers in WTs of Six2 GCE and Cited1 Cre mice. qPCR analysis of early metanephric mesenchyme markers ( Eya1 , Osr1 , Pax2 , Hoxa11 , and Hmga2 ) and renal vesicle markers ( Wnt4 and Jag1 ) in littermate kidneys and tumors from S-Wt1-β-cat S , C-Wt1-β-cat S , and C-Wt1-Igf2 mutants. The x -axis labels indicate wild-type (Wt1 +/fl ) in littermate kidneys and the presence of progenitor-specific Cre alleles ( Six2 GCE or Cited1 Cre ) or Ubiquitous-Cre (U) and the presence of genetic alterations of β-cat S , Wt1 ablation ( Wt1 −/∆ ) or Igf2 upregulation ( H19 +/− m ) in mouse tumors.

Article Snippet: Antibodies used were: WT1 (sc-192, Santa Cruz Biotechnology), Ki67 (ab15580, Abcam), pHH3 (06-570, Upstate Biotechnology), β-catenin (610154, BD Biosciences), Dlk1 (sc-8624, Santa Cruz Biotechnology), Cited1 (9219, Fisher Scientific), Six2 (11562-1-AP, Proteintech), Pax2 (PRB-276P, Covance), NCAM (C9672, Sigma), E-cadherin (3195, Cell Signaling Technology), K-cadherin (ab79005, Abcam), Vimentin (V2258, Sigma), Collagen IV (AB756P, Chemicon), Cyclin D1 (sc-753, Santa Cruz Biotechnology), and C-myc (9E10, Santa Cruz Biotechnology).

Techniques: Expressing

Journal: Molecular Metabolism

Article Title: P2Y2R contributes to the development of diabetic nephropathy by inhibiting autophagy response

doi: 10.1016/j.molmet.2020.101089

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Wilms Tumor-1 (WT1) , Boster Biological Technology , #M00199-1 , Rabbit , 1 μg/ml (IHC).

Techniques:

Journal: Molecular Metabolism

Article Title: P2Y2R contributes to the development of diabetic nephropathy by inhibiting autophagy response

doi: 10.1016/j.molmet.2020.101089

Figure Lengend Snippet: P2Y2R deficiency protects against podocyte loss and glomerular injury in DN. (A) Relative mRNA levels of Nphs1 (nephrin) and Nphs2 (podocin) were determined by real-time PCR analysis (n = 3–5). (B) Localisation of WT1, a podocyte marker, was analysed by immunohistochemistry, and representative images are shown. The number of stained podocytes per glomeruli was counted by using ImageJ software (n = 3). (C) Kidney sections were stained with PAS staining and glomerular morphological changes were scored as described in the method section (n = 3). Data are presented as mean ± SEM. One-way ANOVA was used for statistical analysis followed by Bonferroni's multiple comparisons test. ∗∗∗p < 0.001 vs WT control mice; and ### p < 0.001 vs WT DN mice. Scale bar, 50 μm.

Article Snippet: Wilms Tumor-1 (WT1) , Boster Biological Technology , #M00199-1 , Rabbit , 1 μg/ml (IHC).

Techniques: Real-time Polymerase Chain Reaction, Marker, Immunohistochemistry, Staining, Software, Control

Journal: EMBO Molecular Medicine

Article Title: Inhibition of O‐GlcNAcylation protects from Shiga toxin‐mediated cell injury and lethality in host

doi: 10.15252/emmm.202114678

Figure Lengend Snippet: A Representative immunofluorescence images of the nephron structures within kidney organoids derived from hiPSCs on day 18 of the differentiation; proximal tubules (LTL, green), parietal epithelial cells (PAX8, purple), kidney glomerular podocytes (NPHS1, red), and nephron progenitors (WT1, purple and SIX2, green). Cell nuclei were stained with DAPI. Scale bars: 50 µm. B Representative images showing morphological collapses in iPSC‐derived human kidney organoids following Stx2a (10 ng/ml) treatment for 72 h in the presence or absence OSMI‐1 (10 µM, final). C, D ELISAs were used to analyze the inhibitory effect of OSMI‐1 (10 µM, final) on kidney injury molecule‐1 (KIM‐1) secretion (C) and IL‐8 and CCL‐2 production (D) in culture supernatants of iPSC‐derived human kidney organoids exposed to Stx2a (10 ng/ml) for 72 h ( n = 3 biological replicates). The effects of OSMI‐1 were compared with those of the vehicle (DMSO) controls. E, F Human apoptosis antibody array analysis of multiple proteins using pooled lysates from iPSC‐derived human kidney organoids following Stx2a (10 ng/ml) treatment for 72 h in the presence or absence of OSMI‐1 (10 µM, final). (E) Antibody spots representing signal differences are indicated in red boxes. (F) The graph shows the average of relative spot intensities for each protein compared to those measured in the control in the absence of Stx2a exposure ( n = 2 biological replicates). Dashed line represents the reference point of the fold change. Raw values of fluorescence intensities are provided in the Source Data. Data information: Error bars for bar graphs are presented as mean ± SEM. Statistical analysis was performed using two‐tailed Student’s t ‐test. * P < 0.05; ** P < 0.01; and *** P < 0.001. Source data are available online for this figure.

Article Snippet: WT1 , 1:200 , Cell Signaling Technology 83535 , Alexa Fluor ® 647‐conjugated Donkey Anti‐Rabbit IgG , Jackson ImmunoResearch 711‐605‐152i.

Techniques: Immunofluorescence, Derivative Assay, Staining, Ab Array, Control, Fluorescence, Two Tailed Test

Journal: EMBO Molecular Medicine

Article Title: Inhibition of O‐GlcNAcylation protects from Shiga toxin‐mediated cell injury and lethality in host

doi: 10.15252/emmm.202114678

Figure Lengend Snippet: Primary and secondary antibodies for immunostaining kidney organoids (Subramanian et al , 2019 ).

Article Snippet: WT1 , 1:200 , Cell Signaling Technology 83535 , Alexa Fluor ® 647‐conjugated Donkey Anti‐Rabbit IgG , Jackson ImmunoResearch 711‐605‐152i.

Techniques: Immunostaining, Plasmid Preparation

Journal: Scientific Reports

Article Title: Identification of m6A methyltransferase-related WTAP and ZC3H13 predicts immune infiltrates in glioblastoma

doi: 10.1038/s41598-025-88671-4

Figure Lengend Snippet: Relative expression of genes expression in GBM tissues. Expression levels of WTAP and ZC3H13 genes were compared with those of the control group on GBM tissues, 10 GBM and adjacent tissues, respectively ( A ). The expression levels of siRNA WTAP and overexpression ZC3H13 genes ( B and C ). The expression levels of CD27, CD70, CD80, CD86, ICOS, CTLA4, and LAG3 in siRNA WTAP and overexpression ZC3H13 were demonstrated in Fig. 9D and F, respectively. The result were determined by qRT-PCR. The data are mean ± standard deviation of six replicate experiments. * P < 0.05, **P < 0.01 and ***P < 0.001 compared with the corresponding control values.

Article Snippet: After blocking with 5% nonfat milk, the membranes were immunoblotted with the primary antibodies: WTAP (A04296-2, Boster Biotech, Wuhan, China), CD27 (A01148-2, Boster Biotech), CD70 (A02853-2, Boster Biotech), CD80 (A00196-3, Boster Biotech), CD86(BM4121, Boster Biotech), ICOS (A00291-2, Boster Biotech), CTLA4 (A00020-1, Boster Biotech), LAG3 (M02869-2, Boster Biotech), Beta-actin (BM3873, Boster Biotech).

Techniques: Expressing, Control, Over Expression, Quantitative RT-PCR, Standard Deviation

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: The table presents the quantification of tumor cellularity, PD-L1 or CD274 TPS (Tumor Proportion Score), intensity, LI (lymphocyte infiltration) percentage, and contributions of CD4 and CD8 lymphocytes in LI

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques:

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Expression of immunosuppressive markers correlates to BRAF mutation. (A) The tumor tissues of 19 PTC cases were stained with CD274 antibody, and the expression intensity was scored by a COH pathologist. Grade 1 represents the lowest expression, and grade 3+ represents the highest. (B) The tumor tissue was processed to detect CD73, and the expression intensity was scored where grade 1 represents the lowest expression and grade 3+ represents the highest. ( C ) The images represent the cells positive for cancer cells (TTF-brown), T cells CD8 (blue) and CD4 (pink) in the two representative tumor tissue of BRAF -mutated and WT samples. ( D ) The plot illustrates the percentage of lymphocyte infiltration within the tumor area, as assessed by the pathologist. The lymphocytic infiltration ratio is notably higher for mutant patients (red dots) compared with wild-type patients (blue). Additionally, the percentages of CD4 and CD8 lymphocytes were analyzed. The findings suggest a higher contribution of CD4 lymphocytes compared with CD8, reinforcing the cell type infiltration data. PTC, papillary thyroid cancer; WT, wild-type. * indicates a p value of <0.05

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Expressing, Mutagenesis, Staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: BRAF knockdown induced CD274 expression. (A) The RNA extracted from four PTC cell lines was used to determine the expression of BRAF , CD274, CD73, CD200, CD276, ENTPD1, and CD200. The expression in the mutant was compared with the WT cell type. (B) The immunoblot represents the expression of proteins in the BRAF -mutated and WT cell lines. (C) The bar graph represents the changes in the mRNA expression of CD274 and other immunosuppressive markers on knocking down BRAF . (D) Immunoblot represents the knockdown of BRAF and upregulation of CD274 in the BRAF -mutated cell lines T32 and T85 (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ANOVA test). ANOVA, analysis of variance; PTC, papillary thyroid cancer; WT, wild-type.

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Knockdown, Expressing, Mutagenesis, Western Blot

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: Cell cycle arrest correlates with CD274 expression. (A, B) The T85 cell line was treated with increasing concentration of BRAF inhibitors, and changes in the proliferation were measured using the live cell imaging system. (C, D) Data representing the proliferation changes in the T68 cell lines with respect to BRAF inhibitor treatment. (E) FACS data representing the changes in the cell cycle stages of T68 with respect to BRAF inhibitors where dabrafenib induced stronger cell cycle arrest. (F) The qPCR data represents the changes in the expression of BRAF , CD274 and CD73 in the T68 and T85 cells with respect to the BRAF inhibitor treatment. ( G ) The immunoblot represents the changes in the CD274 and CD73 expression on combining dabrafenib with either binimetinib, temsirolimus, or abemaciclib (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ANOVA test). ANOVA, analysis of variance.

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Expressing, Concentration Assay, Live Cell Imaging, Western Blot

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: BRAF inhibitor-induced transcriptional changes upstream of CD274. (A) Cartoon showing the design of CD274-promoter-luciferase constructs used for determining the changes in transcriptional activity postdrug treatment. (B) The box plot representing the luciferase activity determined in the cells post 48 hours of transfection, where 2 Kb promoter sequence correlated with maximum activity. (C) Luciferase activity measured in the cells post 48 hours of BRAF or KRAS knockdown. ( D ) The bar graph represents the difference in the promoter activity between the T85 and T32 cell lines. (E, F) Luciferase (Promoter) activity correlated with increased BRAF inhibitor concentration in the T68 and T85 cells. (****p<0.0001, ANOVA test). ANOVA, analysis of variance.

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Luciferase, Construct, Activity Assay, Transfection, Sequencing, Knockdown, Concentration Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: The drug combination effect on CD274 expression. (A, B) The bar graph represents the percentage change in the growth of T68 or T85 cells on binimetinib, abemaciclib or temsirolimus exposure. The immunoblot shows the change in the expression of CD274 and CD73 on drug treatment. (C) The bar graph represents the effect of drug combinations on the cell viability of T85 cells as measured by CCK8 assay. (D–F) The immunoblot represents the increase in the expression of CD274 and CD73 expression on combining dabrafenib with either binimetinib or temsirolimus, or abemaciclib (*****p<0.0001, ANOVA test). ANOVA, analysis of variance.

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Exploring markers of immunoresponsiveness in papillary thyroid carcinoma and future treatment strategies

doi: 10.1136/jitc-2023-008505

Figure Lengend Snippet: The synergistic effect of BRAF and checkpoint inhibitors in immune cell-mediated killing. (A) The line graph represents the growth kinetics of T85 cells when cultured in combination with activated CD8 cells. The bar graph represents the comparison in growth between monoculture and cocultured T85 cells at 0 and 72 hours time points. (B) The growth kinetics of T32 cells when cultured in combination with activated CD8 cells. (C) The images represent the effect of dabrafenib, CD8, atezolizumab, and IL-2 on the T85 (GFP) cell growth. (D) The growth kinetics of T85 cells when cultured in combination with activated CD8 cells, IL-2, and atezolizumab. (E) The growth kinetics of T85 cells, when cultured in combination with dabrafenib, activated CD8 cells, IL-2 and atezolizumab. (F) The images represent the effect of CD8, atezolizumab, and IL-2 on the T85 (GFP) derived spheroids. Loss of GFP represents a loss of spheroid viability. (G) The bar graph represents the changes in the spheroid viability when cocultured with CD8, IL-2, and atezolizumab. (H) The changes in the spheroid viability when cocultured with dabrafenib, CD8, IL-2, and atezolizumab. (I) The immunoblot represents the changes in the CD274 expression in T85 cells on exposure to IL-2 and dabrafenib n monoculture or coculture. **** indicates P value < 0.001.

Article Snippet: Likewise, the pGL3 plasmid harboring the WT or mutant UTR region of the CD274 gene were purchased from Addgene (#107009, #107010).

Techniques: Cell Culture, Comparison, Derivative Assay, Western Blot, Expressing

Journal: Biochemical Journal

Article Title: WT1 and its transcriptional cofactor BASP1 redirect the differentiation pathway of an established blood cell line

doi: 10.1042/bj20101734

Figure Lengend Snippet: Figure 2 Expression of BASP1 in K562 cells leads to the general down-regulation of WT1 target genes

Article Snippet: Control siRNAs (small interfering RNAs) used were from Ambion (AM4611 and AM4635) and WT1 siRNAs were from Santa Cruz Biotechnology (sc-36846) and Ambion (s14912).

Techniques: Expressing

Journal: Biochemical Journal

Article Title: WT1 and its transcriptional cofactor BASP1 redirect the differentiation pathway of an established blood cell line

doi: 10.1042/bj20101734

Figure Lengend Snippet: Figure 5 WT1 is required for the altered PMA-dependent differentiation programme of B-K562 cells

Article Snippet: Control siRNAs (small interfering RNAs) used were from Ambion (AM4611 and AM4635) and WT1 siRNAs were from Santa Cruz Biotechnology (sc-36846) and Ambion (s14912).

Techniques: