Journal: Molecular Cancer

Article Title: PIK3CA alterations in Middle Eastern ovarian cancers

doi: 10.1186/1476-4598-8-51

Figure Lengend Snippet: Antibodies used for tissue microarray Immunohistochemical analysis

Article Snippet: p-AKT (Cytoplasmic & Nuclear) , 75/144 52.1 , Ser473 , Cell Signaling , Mouse Mono-clonal , Predilute , pH 9, microwave , Survival Marker; Signal Stain IHC detection Kit.

Techniques: Microarray, Immunohistochemical staining, Marker, Staining

Journal: Frontiers in Immunology

Article Title: Attenuated Salmonella potentiate PD-L1 blockade immunotherapy in a preclinical model of colorectal cancer

doi: 10.3389/fimmu.2022.1017780

Figure Lengend Snippet: PD-1 and PD-L1 are expressed in MC38 tumors. Representative flow cytometric dot plots showing the expression of PD-L1 (A, C) and PD-1 (B, D) on MC38 tumor cells grown in vitro (A, B) or on single cell suspension of dissociated tumor tissues excised from mice 21 post subcutaneous implantation (C, D) . Immunohistochemical staining was also used to illustrate the expression of PD-L1 (E) and PD-1 (F) on MC38 tumor tissue sections. Magnification 400×. Scale bar 20 μm.

Article Snippet: All immunohistochemical studies were done using the described protocol except for cleaved caspase-3 staining which was done following the protocol of IHC Detection Kit of the manufacturer (Cell Signaling Technology; #12692).

Techniques: Expressing, In Vitro, Immunohistochemical staining, Staining

Journal: Frontiers in Immunology

Article Title: Attenuated Salmonella potentiate PD-L1 blockade immunotherapy in a preclinical model of colorectal cancer

doi: 10.3389/fimmu.2022.1017780

Figure Lengend Snippet: Combination treatment increases MC38 tumor infiltration by CD4 + and CD8 + T cells and the expression of their effector molecules. MC38 tumor tissues were resected from non-treated, αPD-L1 and combination-treated mice on day 21 post implantation for further analyses using immunohistochemistry, flow cytometry and qRT-PCR. Representative immunohistochemical images for CD4 (A) and CD8 (C) in tumor tissues are presented for the control and combination-treated groups. Magnification 400×. Scale bar 20 μm. CD4 + (B) and CD8 + (D) cells were quantified in 15 HPF for control, αPD-L1 and combination-treated groups. Each data point represents the average of positive cells/HPF from a single mouse, pooled from 2 independent experiments. Representative flow cytometric dot plots and the combined result analyses for the percentages of CD4 + (E, F) and CD8 + (E, G) cells in MC38 tumors are illustrated. RNA was extracted from total MC38 tumor tissues and gene expression levels were determined using qRT-PCR. The effect of combination treatment on the expression levels of CXCL9 (H) , CXCL10 (I) , IFN-γ (J) and granzyme B (K) was assessed. Representative images for granzyme B staining in tumor tissues are presented for each group (L) . Graph depicts the number of cells/HPF (M) . Magnification 400×. Scale bar 20 μm. Each data point represents a single mouse pooled from two independent experiments. Asterisks denote statistically significant differences from control group, * (P ≤ 0.05), ** (P ≤ 0.01), *** (P ≤ 0.001) and **** (P ≤ 0.0001).

Article Snippet: All immunohistochemical studies were done using the described protocol except for cleaved caspase-3 staining which was done following the protocol of IHC Detection Kit of the manufacturer (Cell Signaling Technology; #12692).

Techniques: Expressing, Immunohistochemistry, Flow Cytometry, Quantitative RT-PCR, Immunohistochemical staining, Staining

Journal: Frontiers in Immunology

Article Title: Attenuated Salmonella potentiate PD-L1 blockade immunotherapy in a preclinical model of colorectal cancer

doi: 10.3389/fimmu.2022.1017780

Figure Lengend Snippet: Combination treatment of Salmonella and αPD-L1 induces apoptosis more efficiently than monotherapy. Representative immunohistochemical images for cleaved caspase-3 in tumor tissues are presented for control, Salmonella , αPD-L1 and combination-treated groups (A) . Magnification 400×. Scale bar 20 μm. Cleaved caspase 3 + cells were quantified in 15 HPF for the different groups (B) . Each data point represents the average of positive cells from a single mouse, pooled from 2 independent experiments for all groups except for the control group, from three independent experiments. Asterisks denote statistically significant differences from control group, * (P ≤ 0.05), ** (P ≤ 0.01), *** (P ≤ 0.001) and ns (no statistical significance, ≥ 0.05).

Article Snippet: All immunohistochemical studies were done using the described protocol except for cleaved caspase-3 staining which was done following the protocol of IHC Detection Kit of the manufacturer (Cell Signaling Technology; #12692).

Techniques: Immunohistochemical staining

Journal: PLoS ONE

Article Title: The Antidiabetic Drug Ciglitazone Induces High Grade Bladder Cancer Cells Apoptosis through the Up-Regulation of TRAIL

doi: 10.1371/journal.pone.0028354

Figure Lengend Snippet: Mice were inoculated subcutaneously with exponentially growing T24 and RT4 bladder cancer cells (5×10 6 cells). When tumor size reached 40 mm 3 in volume, mice were randomly divided into control and treated groups (n = 10). Intraperitoneal injections of ciglitazone were weekly administered at a dose of 15 mg/kg during three weeks. Control animals received only saline vehicle following an identical schedule. (A) The growth tumor curves were determined by measuring the tumor volume. * P <0.05, significant differences compared with untreated animals with the use of two-way ANOVA test (evaluation of the tumor volume development over time) ; ** P <0.05, significant differences between control and treated mice at each post-graft time with the use of two-tailed unpaired Student's t test. (B) Immunohistochemical staining of representative paraffin-embedded sections of tumors from untreated or ciglitazone-treated mice. Sections were fixed and stained for Ki-67 and active caspase 3. Each panel is representative of 5 sections for each of ten tumors from control and ciglitazone-treated mice. Original magnification, ×20.

Article Snippet: Activated caspase 3 was detected with Apoptosis marker: SignalStain cleaved Caspase 3 (Asp175) IHC Detection kit (Cell Signaling).

Techniques: Two Tailed Test, Immunohistochemical staining, Staining

Journal: PLoS ONE

Article Title: Antitumor Activity of Noscapine in Combination with Doxorubicin in Triple Negative Breast Cancer

doi: 10.1371/journal.pone.0017733

Figure Lengend Snippet: Tumor sections were stained using the DeadEnd colorimetric kit and cleaved caspase-3 (Asp 175) IHC kit for TUNEL assay and cleaved caspase 3 expression as described in materials and methods respectively. The apoptotic tumor cells are stained brown. Percentages of TUNEL-positive and cleaved caspase 3-positive cells were quantitated by counting 100 cells from 6 random microscopic fields. Data are expressed as mean+SD (N = 6). One-way ANOVA followed by post Tukey test was used for statistical analysis to compare control and treated groups. P <0.01 (*, significantly different from untreated controls; ** , significantly different from Noscapine and Doxorubicin single treatments). Original magnification ×40 (Micron bar = 100 µm).

Article Snippet: Tumor tissue sections prepared from formalin-fixed, paraffin-embedded tumor tissues were used for IHC studies according to the protocol specified in the SignalStain™ Cleaved Caspase-3 (Asp 175) IHC kit (Cell Signaling, Beverly, MA).

Techniques: Staining, TUNEL Assay, Expressing

Journal: PLoS ONE

Article Title: Modulation of Glucose Transporter 1 (GLUT1) Expression Levels Alters Mouse Mammary Tumor Cell Growth In Vitro and In Vivo

doi: 10.1371/journal.pone.0023205

Figure Lengend Snippet: A. 0.5 million 78617GL cells expressing control shRNA (C) or GLUT1 shRNA (G1) were injected into contralateral #4 mammary fat pads of athymic nude mice. Bioluminescence from the labeled tumor cells was detected on days 2, 4, 6 and 8 after implantation. The abdominal region heat map depicting luciferase activity of a representative mouse is pictured. B. The average bioluminescence on days 2, 4, 6 and 8 normalized to the day 2 bioluminescence +/− SEM for five mice is presented with the black diamonds representing shCTRL tumors and grey squares representing shGLUT1 tumors. C. Expression of GLUT1 and β-actin in lysates of three tumor pairs evaluated by immunoblot analysis. D. GLUT1 expression evaluated by IHC (with hematoxylin counterstain) in two tumor pairs. E. Representative low power photomicrographs of two pairs of tumor sections immunostained for cleaved caspase 3. F–G. Representative high power photomicrographs of a tumor pair immunostained for BrdU with hematoxylin counterstain ( F ) which is quantified ( G ).

Article Snippet: Detection of apoptotic cells was performed similarly using SignalStain cleaved caspase-3 IHC detection kit (Cell Signaling Technologies, Beverly, MA) with pre-diluted rabbit anti-cleaved capsase-3 antibody.

Techniques: Expressing, shRNA, Injection, Labeling, Luciferase, Activity Assay, Western Blot

Journal: PLoS ONE

Article Title: Modulation of Glucose Transporter 1 (GLUT1) Expression Levels Alters Mouse Mammary Tumor Cell Growth In Vitro and In Vivo

doi: 10.1371/journal.pone.0023205

Figure Lengend Snippet: A. 0.5 million G1fPt cells infected two weeks prior with adenovirus expressing GFP or Cre recombinase were injected into contralateral #4 mammary fat pads of athymic nude mice. Bioluminescence from the labeled tumor cells was detected on days 12, 15 and 20 after implantation. The abdominal region heat map depicting luciferase activity of a representative mouse is pictured. B. The average bioluminescence on days 12, 15 and 20 +/− SEM for eight mice is presented with green diamonds representing “GFP” tumors and light blue squares representing “Cre” tumors. C. Expression of GLUT1 and β-actin in lysates of two tumor pairs evaluated by immunoblot analysis. D. GLUT1 expression evaluated by IHC (with hematoxylin counterstain) in two tumor pairs. E. Representative low power photomicrographs of two pairs of tumor sections immunostained for cleaved caspase 3. F–G. Representative high power photomicrographs of a tumor pair immunostained for Ki67 with hematoxylin counterstain ( F ) which is quantified ( G ).

Article Snippet: Detection of apoptotic cells was performed similarly using SignalStain cleaved caspase-3 IHC detection kit (Cell Signaling Technologies, Beverly, MA) with pre-diluted rabbit anti-cleaved capsase-3 antibody.

Techniques: Infection, Expressing, Injection, Labeling, Luciferase, Activity Assay, Western Blot

Journal: PLoS ONE

Article Title: Modulation of Glucose Transporter 1 (GLUT1) Expression Levels Alters Mouse Mammary Tumor Cell Growth In Vitro and In Vivo

doi: 10.1371/journal.pone.0023205

Figure Lengend Snippet: A. 0.4 million 85815GL cells expressing control vector (V) or GLUT1 (G1) were injected into contralateral #4 mammary fat pads of athymic nude mice. Bioluminescence from the labeled tumor cells was detected on days 3, 6, 9, 12 and 14 after implantation. B. The bioluminescence on days 3, 6, 9, 12 and 14 normalized to the day 3 value was averaged for all five mice and is presented +/−SEM. C. GLUT1 and β-actin expression evaluated in lysates of three tumor pairs by immunoblot analysis. D. GLUT1 expression evaluated by IHC in two tumor pairs. E. Low power photomicrographs of a pair of tumor sections derived from a vector control tumor (top) and a tumor overexpressing GLUT1 (bottom) immunostained for cleaved caspase 3 (left). High power photomicrographs of a pair of tumor sections immunostained for cleaved caspase 3 (middle), which are converted to binary pictures (right). F. Quantification of the number of cleaved caspase 3 positive pixels in five high power field “hot spots” in four tumors of each group. G–H. Representative high power photomicrographs of tumor sections immunostained for BrdU with hematoxylin counterstain ( G ) which is quantified ( H ).

Article Snippet: Detection of apoptotic cells was performed similarly using SignalStain cleaved caspase-3 IHC detection kit (Cell Signaling Technologies, Beverly, MA) with pre-diluted rabbit anti-cleaved capsase-3 antibody.

Techniques: Expressing, Plasmid Preparation, Injection, Labeling, Western Blot, Derivative Assay