Journal: bioRxiv

Article Title: Targeting wild-type IDH1 enhances chemosensitivity in pancreatic cancer

doi: 10.1101/2023.03.29.534596

Figure Lengend Snippet: A] Tumor growth curves of PDX (TM01212) in nude mice treated with vehicle, AG-120, 5-FU, or AG-120 and 5-FU for a total of 37 days. Treatment was started when tumors reached 100-120mm 3 in size. Tumor volumes were calculated twice per week using calipers (n=5 per group). B] Average tumor volume at the end of the experiment (day 68) (n=5 tumors per group). C] Representative image of in vivo and excised PDX tumors at the end of the experiment. D] Average tumor weights (mg) of PDX tumors in each group (n=5 per group). E] Body weights of nude mice bearing PDX (n=5 per group). The paraffin-embedded tissue sections derived from nude mice bearing TM01212 PDX. F] Cell proliferation in PDX tumors was assessed by nuclear immunolabeling by Ki-67. Scale bar, 100 μm. G] Apoptosis was assessed in PDX tumors with labeled cleaved caspase-3. Scale bar, 100 μm. Each data point represents the mean ± SEM of at least three independent experiments. N.S., nonsignificant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The primary antibodies were diluted (SignalStain Antibody Diluent; 8112) and incubated overnight at 4 °C using the following antibodies: Ki67 (SP6, BioCare Medical) and cleaved caspase 3 (Cell Signaling Technology; 9579).

Techniques: In Vivo, Derivative Assay, Immunolabeling, Labeling

Journal: Cell Communication and Signaling : CCS

Article Title: Estrogen downregulates CD73/adenosine axis hyperactivity via adaptive modulation PI3K/Akt signaling to prevent myocarditis and arrhythmias during chronic catecholamines stress

doi: 10.1186/s12964-023-01052-0

Figure Lengend Snippet: Antibody used in the study

Article Snippet: 8 , Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb , Cell Signalling Technology , #9579.

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Estrogen downregulates CD73/adenosine axis hyperactivity via adaptive modulation PI3K/Akt signaling to prevent myocarditis and arrhythmias during chronic catecholamines stress

doi: 10.1186/s12964-023-01052-0

Figure Lengend Snippet: In vivo results of estrogenic adaptive regulation of CD73/adenosine axis via PI3K/Akt pathways during CCS. A – D Representative immunoblots and graphical presentations of cleaved caspase 3, PI3K, p-PI3K and Akt, p-Akt, were assessed from Mice. S = Sham; S + I = Sham + ISO; O = OVX; O + I = OVX + ISO; O + E, OVX + E2; O + E + I = OVX + ISO + E2. E – H In vitro results of inhibition of PI3Kby LY294002. Representative immunoblots and graphical presentations of, PI3K, p-PI3K, Akt, p-Akt and CD73. I , J Representative immunofluorescence and plotted average values of CC3 expression. Data are presented as mean ± SEM; *** p < 0.001; ** p < 0.01; * p < 0.05

Article Snippet: 8 , Cleaved Caspase-3 (Asp175) (D3E9) Rabbit mAb , Cell Signalling Technology , #9579.

Techniques: In Vivo, Western Blot, In Vitro, Inhibition, Immunofluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Secondary Degeneration of Oligodendrocyte Precursor Cells Occurs as Early as 24 h after Optic Nerve Injury in Rats

doi: 10.3390/ijms24043463

Figure Lengend Snippet: Effect of injury on apoptosis within the ventral optic nerve. ( A ) The density of Cleaved Caspase3+ cells was quantified in the ventral optic nerve following partial optic nerve transection or sham injury. Graphs display individual data points overlaid on a bar displaying the mean ± SEM. n = 9 rats per group. Statistical analysis by t -test. ( B ) The relative densities of Cleaved Caspase3+ 8OHDG+ and Cleaved Caspase3+ 8OHDG− cells were quantified. Statistical analysis by two-way ANOVA and Tukey post hoc tests. Graphs display individual data points overlaid on a bar displaying the mean ± SEM. n = 9 rats per group. ( C ) The area of IgG immunointensity was correlated to the density of Cleaved Caspase3+ cells using Spearman’s correlation, with the r s value and corresponding p -value displayed on the graph. The mean area of IgG immunointensity was plotted on a log scale to best illustrate the overall relationship between IgG and Cleaved Caspase3+ on the scatterplot. Each data point on the graph represents an individual animal. n = 8–9 rats per group. ( D ) Representative image of both a Cleaved Caspase3+ 8OHDG+ cell (indicated by >>) and a Cleaved Caspase3+ 8OHDG− cell (indicated by >) is shown, scale bar = 25 μm. No outliers were removed for any outcome measure. Significant differences are indicated by ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Primary antibodies used recognized: 8OHDG (1:250, 4 µg/mL, mouse, Abcam, ab62623, GR3284216-13), rat Immunoglobulin G (IgG, 1:150, 10 µg/mL, goat, BA-9400, ZG0108, Vector Laboratories, Burlingame, CA, USA), NG2 (1:50, 20 µg/mL, rabbit, AB5320B, 3218879, Merck), Cleaved Caspase3 (Asp175, 1:150; rabbit, D3E9, #9579, Lot 1, Cell Signaling Technology, Danvers, MA, USA) and PDGFRα (1:250, 4 µg/mL, rabbit, PA516571, VJ2870528A, ThermoFisher, Waltham, MA, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Secondary Degeneration of Oligodendrocyte Precursor Cells Occurs as Early as 24 h after Optic Nerve Injury in Rats

doi: 10.3390/ijms24043463

Figure Lengend Snippet: Effect of injury on apoptosis and oxidative DNA damage of OPCs. ( A ) The relative densities of Cleaved Caspase3+ PDGFRα+ and Cleaved Caspase3+ PDGFRα− OPCs were quantified. Statistical analysis by two-way ANOVA and Tukey post hoc tests. ( B ) The relative densities of Cleaved Caspase3+ cells colocalized with PDGFRα and 8OHDG were quantified. Statistical analysis by three-way ANOVA with Tukey post hoc tests. Graphs display individual data points overlaid on a bar displaying the mean ± SEM. n = 9 rats per group. ( C ) Representative image illustrating Cleaved Caspase3+ PDGFRα+ 8OHDG+ cells (indicated by >>), Cleaved Caspase3+ PDGFRα+ 8OHDG− cells (indicated by >|) and Cleaved Caspase3+ PDGFRα− 8OHDG+ cells (indicated by >) is shown. Cleaved Caspase3+ PDGFRα− 8OHDG− cells were not observed in the ventral nerve. Scale bar = 25μm. No outliers were removed for any outcome measure. Significant differences are indicated by * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001.

Article Snippet: Primary antibodies used recognized: 8OHDG (1:250, 4 µg/mL, mouse, Abcam, ab62623, GR3284216-13), rat Immunoglobulin G (IgG, 1:150, 10 µg/mL, goat, BA-9400, ZG0108, Vector Laboratories, Burlingame, CA, USA), NG2 (1:50, 20 µg/mL, rabbit, AB5320B, 3218879, Merck), Cleaved Caspase3 (Asp175, 1:150; rabbit, D3E9, #9579, Lot 1, Cell Signaling Technology, Danvers, MA, USA) and PDGFRα (1:250, 4 µg/mL, rabbit, PA516571, VJ2870528A, ThermoFisher, Waltham, MA, USA).

Techniques:

Journal: International Journal of Experimental Pathology

Article Title: Apoptosis versus necrosis in tubal ectopic pregnancies following Methotrexate

doi: 10.1111/iep.12465

Figure Lengend Snippet: (A) Fallopian Tube, H&E Staining. BC, Blood Clot; MS, Muscular Wall; PL, Plica. Magnification 100×. (B) Villi in Fallopian Tube. The Villi is surrounded by RBC. The outer layer is composed of Syncytiotrophoblast. The inner layer is composed of Cytotrophoblast. The stroma of the Villi contains Hofbauer cells (Macrophages). Haematoxylin and Eosin Stain, Magnification 100×. (C) Villi Fallopian Tube, Same area as Figure 2, Cleaved Caspase – 3 Staining. Red Arrow – Apoptotic Epithelial Cell, Magnification 100×.

Article Snippet: The specimens were put on slides and dyed using the Cleaved Caspase‐3 (Asp175) Rabbit monoclonal antibody (Cleaved Caspase‐3 antibody, #9579; Cell Signaling Technology).

Techniques: Staining, H&E Stain

Journal: Frontiers in Endocrinology

Article Title: Corticosterone induces obesity partly via promoting intestinal cell proliferation and survival

doi: 10.3389/fendo.2022.1052487

Figure Lengend Snippet: CORT increased intestinal villus length by breaking the balance between proliferation and death of intestinal cells. (A) Hematoxylin and eosin (H&E)-staining for duodenum sections. (B) Quantitative analysis of villi length for duodenum from Vehicle- or CORT-treated mice. (C) Immunofluorescence staining of EdU for the duodenum sections from Vehicle- or CORT-treated mice. (D) Quantification of the progression of EdU-labelled cells along the duodenum villi from Vehicle- or CORT-treated mice. (E) Immunofluorescence staining of Ki67 for the duodenum sections from Vehicle- or CORT-treated mice. (F) Quantification of the duodenal crypt depth indicated by Ki67-staining from Vehicle- or CORT-treated mice. (G) Immunofluorescence staining of TUNEL for the duodenum sections from Vehicle- or CORT-treated mice. (H) Quantification of TUNEL-positive cells of duodenal crypt from Vehicle- or CORT-treated mice. (I) Immunofluorescence staining of CC3 for the duodenum sections from Vehicle- or CORT-treated mice. (J) Quantification of CC3-positive cells of duodenal crypt from Vehicle- or CORT-treated mice. Data are expressed as mean ± SEM, *p < 0.05; **p < 0.01. CC3: Cleaved Caspase 3. Veh: vehicle-treated group; CORT: corticosterone-treated group.

Article Snippet: Primary antibodies used for immunofluorescence: Ki67 (rabbit, Abcam ab16667, dilution 1:200), Cleaved Caspase 3 (CC3) (rabbit, Cell Signaling Technologies 9579S, dilution 1:200).

Techniques: Staining, Immunofluorescence, TUNEL Assay