Journal: Journal of colloid and interface science

Article Title: Cubosomes functionalized with antibodies as a potential strategy for the treatment of HER2-positive breast cancer.

doi: 10.1016/j.jcis.2024.06.091

Figure Lengend Snippet: Fig. 1. Proposed nanotechnological platform for targeting HER2-positive breast cancer cells. (A) Design and components of the panobinostat-loaded cubosomes functionalized with trastuzumab. (B) Panobinostat-loaded immunocubosomes administrated intravenously to treat HER2-positive breast cancer. (C) Cellular uptake, (C1) panobinostat-loaded immunocubosomes internalization by breast cancer cell line expressing HER2 (SKBR3), (C2) interaction between HER2 and trastuzumab, (C3) steps of endocytosis, and (C4) cell death.

Article Snippet: For cells, the L929 mouse fibroblast and 4T1 breast cancer were a generous gift from Prof. Dr. Jean-Christophe Leroux’s laboratory, and the SKBR3 breast cancer was purchased from DSMZ, Germany.

Techniques: Expressing

Journal: Journal of colloid and interface science

Article Title: Cubosomes functionalized with antibodies as a potential strategy for the treatment of HER2-positive breast cancer.

doi: 10.1016/j.jcis.2024.06.091

Figure Lengend Snippet: Fig. 4. Cellular uptake and cytotoxicity assessment of free Panobinostat (Free Pa), blank cubosomes (Cub), panobinostat-loaded cubosomes (CubPa) and panobinostat-loaded immunocubosomes (ICubPa) by L929, 4T1, and SKBR3 cell lines. (A) Fluorescence microscopy images of cellular uptake of Dio-loaded cubo somes and Dio-loaded immunocubosomes by L929, 4T1, and SKBR3 cell lines. The first line represents DAPI staining (nuclei, blue), the second line Dio staining (cytoplasm, green), and the third line the overlay of DAPI and Dio staining. Scale bar 50 μm. (B) Cell viability of L929, 4T1, and SKBR3 cells incubated with free panobinostat, blank cubosomes, and panobinostat-loaded cubosomes for 24 h. (C) Cell viability of L929, 4T1, and SKBR3 cells incubated with free panobinostat, panobinostat-loaded cubosomes, and panobinostat-loaded immunocubosomes for 24 h. (D) Analysis of the effectiveness of treatments by observing cell viability and cell death of SKBR3 cell incubated with free panobinostat, panobinostat-loaded cubosomes, and panobinostat-loaded imunocubosomes for 24 h. (E) Analysis of the selectivity of antibody therapy by observing cell viability and cell death of L929 and SKBR3 cells incubated with panobinostat-loaded imunocubosomes for 24 h. Data are shown as mean ± SD of at least six experiments. *Denotes statistical differences between groups, two-way ANOVA, Tukey *p < 0.05.

Article Snippet: For cells, the L929 mouse fibroblast and 4T1 breast cancer were a generous gift from Prof. Dr. Jean-Christophe Leroux’s laboratory, and the SKBR3 breast cancer was purchased from DSMZ, Germany.

Techniques: Fluorescence, Microscopy, Staining, Incubation

Journal: Plants

Article Title: Screening for Selective Anticancer Activity of 65 Extracts of Plants Collected in Western Andalusia, Spain

doi: 10.3390/plants10102193

Figure Lengend Snippet: Cytotoxic activity of Tetraclinis articulata (Vahl) Mast. ( 58 ) against 14 cancer cell lines from a variety of tissues.

Article Snippet: HNO97 (human tongue cancer cells), A64-CLS (human submaxillary gland adenoma cells), AN3Ca (human endometrial adenocarcinoma cells), Sk-OV-3 (human ovarian cancer cells), KATO III (human gastric cancer cells), Sk-Br-3 (HER2-positive breast cancer cells), T24 (human bladder cancer cells), Calu-1 (human squamous lung cancer cells), and MeWo (human BRAF wild-type melanoma cells) were purchased from Cell Lines Service (CLS).

Techniques: Activity Assay

Journal: Nucleic Acids Research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Local RNA in situ hybridization. ( A ) Schematic overview of a microfluidic implementation of chromogenic RNA in situ hybridization. The initial preparation of the tissue section including paraffin removal (1), heat (2) and protease (3) pretreatment, as well as fixation was performed globally. The primary probe was then hybridized locally to the RNA of interest (4). In this case the detection of HER2 is shown with internal negative ( dapB ) and positive ( ActB ) controls. Finally, the amplification of the signal using preamplifier and amplifier strands (5), the binding of enzyme (6), and the enzymatic reactions (7) were performed globally on the whole slide. ( B ) Fluorescence images of RNA-ISH experiments detecting the gene expression of HER2 in cell pellet sections of the breast cancer cell lines MCF7 (left), SKBR3 (center) and BT474 (right). Probes directed against bacterial dapB and ActB were used as negative and positive controls, respectively. Scale bar: 100 μm.

Article Snippet: FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: RNA In Situ Hybridization, Amplification, Binding Assay, Fluorescence, Expressing

Journal: Nucleic Acids Research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Detection of HER2 expression status by RNA in situ hybridization using internal positive and negative controls. ( A ) Fluorescence images of the signals detected for HER2 , dapB , and ActB in a mammary carcinoma section. Scale bar: 100 μm. ( B ) Quantitative analysis of the fluorescence intensity of FISH signals detected for HER2 , dapB , and ActB integrated per sectioned cell for FFPE sections of the breast cancer cell lines MCF7, SKBR3, and BT474 (see Figure ) as well as the mammary carcinoma from (A). 100 ms excitation.

Article Snippet: FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: Expressing, RNA In Situ Hybridization, Fluorescence

Journal: Nucleic Acids Research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Single color multiplexed RNA-ISH for the detection of breast cancer biomarkers. ( A ) The primary probes for ER , PgR , and HER2 were delivered to spatially distinct regions of a mammary carcinoma section using a microfluidic chip (schematic in upper left corner). Fluorescence images of the signal detected for the breast cancer biomarkers ER , PgR and HER2 in a mammary carcinoma section. Scale bar: 100 μm. ( B ) Fluorescence intensity of FISH signals detected for ER , PgR and HER2 integrated per sectioned cell for FFPE sections of the breast cancer cell lines MCF7, SKBR3 and BT474 (see ) as well as the mammary carcinoma from (A). 500 ms excitation.

Article Snippet: FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: Fluorescence

Journal: Biology Open

Article Title: Meningeal retinoic acid contributes to neocortical lamination and radial migration during mouse brain development

doi: 10.1242/bio.021063

Figure Lengend Snippet: Postnatal Raldh2cKO mice have decreased neuron numbers in upper cortical layers. (A-J′) Labellings on brain sections from P4 control and Raldh2cKO animals for DAPI (A-B′) and immunolabellings for Cux1 (C-D′), Brn2 (E-F′) or RORβ (I-J′) at rostral (A-J) levels and caudal (A′-J′) levels. (G-H′) Overlay of Cux1 and Brn2 signals. White boxes in A-B′ show the areas selected for the higher magnification pictures of the Cux1, Brn2 and RORβ immunolabellings, and in C-J′ the areas used for cell counts. (K) Quantification of Cux1-positive cells; rostrally: 388.4±17.76 for control and 333.26±12.68 for Raldh2cKO; caudally: 347.6±18.68 for control and 273.73±15.50 for Raldh2cKO. (L) Quantification of Brn2-positive cells; rostrally: 248.26±12.60 for control and 287.2±8.22 for Raldh2cKO; caudally: 221.06±18.91 for control and 243.4±17.53 for Raldh2cKO. (M) Quantification of Cux1-positive/Brn2-negative cells; rostrally: 67.25±5.18 for control and 45.58±1.14 for Raldh2cKO; caudally: 63.66±7.25 for control and 33.83±4.27 for Raldh2cKO. (N) Quantification of RORβ-positive cells; rostrally: 97.33±5.95 for control and 76.8±4.11 for Raldh2cKO; caudally: 84.53±6.73 for control and 59.93±5.71 for Raldh2cKO. Data presented as mean±s.e.m.; n =5 brains; * P <0.05; ns, not significant by two-tailed Student's t -test. Scale bars: 400 μm (A-B′), 100 μm (C-J′).

Article Snippet: After antigen unmasking in citrate buffer (0.01 M, pH 6) during 15 min in a microwave oven, slides were blocked with 5% donkey serum, 0.1% Triton X-100 in phosphate-buffered saline (PBS) and incubated overnight with the following primary antibodies: bromodeoxyuridine (BrdU) (1:500, AbD Serotec #OBT0030G), Ki67 (1:300, Novocastra #NCL-KI67P), phospho-histone H3 (1:500, Upstate #05-806); Pax6 (1:300, Covance #PRB-278P); Tbr2 (1:300, eBioscience #14-4875); βIII-tubulin/Tuj1 (1:200, Covance #MMS-435P-100); Tbr1 (1:100, Abcam #ab31940); Ctip2 (1:500, Abcam #ab18465); Cux1 (1:100, Santa Cruz Biotechnologies #sc13024); cleaved caspase-3 (1:200, R&D Systems #NB100-56113); Brn2 (1:1000 Santa Cruz Biotechnologies #sc6029X), RORβ (1:500, Santa Cruz Biotechnologies #sc21354); Calretinin (1:2000, Swant #7699/4); Raldh2 (1:75, Santa Cruz Biotechnologies #sc22591); nestin (1:100; Developmental Studies Hybridoma Bank #rat-401); laminin (1:500, Sigma #L9393).

Techniques: Control, Two Tailed Test