antibody based protein array  (R&D Systems)

Journal: Clinical & Experimental Metastasis

Article Title: A CSF-1R inhibitor both prevents and treats triple-negative breast cancer brain metastases in hematogenous preclinical models

doi: 10.1007/s10585-025-10366-x

Figure Lengend Snippet: CSF-1R suppression alters microglial cytokine secretion patterns with effects on astrocytes and tumor cells. A Mouse cytokine antibody array analysis of EOC2-derived conditioned medium treated with 100 nM BLZ945 or DMSO for 72 h. The images of cytokines with marked GM-CSF, IL-1Rα, IL-6 and TNFα are shown in the left panel. The pixel density of the above cytokines relative to the internal control is shown by the graphs in the right panel. B RT-PCR analysis of GM-CSF , IL-1Rα , IL-6 , and TNFα was performed on fixed, frozen specimens from vehicle control and BLZ945 treatment settings of the 4T1-BR5 TNBC brain metastasis model mice on Days -14, 3, and 10. Values represent the fold change ± SD relative to vehicle. Each dot represents one mouse, and the line designates the group median. C 4T1-BR5 and 231-BR cells were allowed to invade through Matrigel for 12 h in the presence of control or BLZ945 treated EOC2-derived conditioned medium. Invaded cells were visualized by Hema 3 staining and counted using ImageJ software. D and E EOC2 conditioned medium promotes the growth of 4T1-BR5 and 231-BR cells under co-culture conditions. 4T1-BR5 and 231-BR cells were labelled with mCherry and EGFP, respectively. Brain trophic cancer cell numbers are quantified and calculated by flow cytometry based on the ratio between cancer cells and quantitative beads. The effect of astrocytes alone on cancer cell growth was measured in low serum medium under co-culture conditions. n ≥ 3 biologically independent experiments. *, p < 0.05 vs DMSO control; n.s., not significant by the Mann–Whitney test. F Schematic representation of microglial action in breast cancer brain metastasis. CSF-1R + microglia/macrophages regulate the secretion of cytokines that influence the behavior of astrocytes and brain metastatic (BrM) cells within the microenvironment

Article Snippet: The mouse cytokine antibody array (#ARY006, R&D Systems) was used according to the manufacturer's instructions and analyzed using the ImageJ software.

Techniques: Ab Array, Derivative Assay, Control, Reverse Transcription Polymerase Chain Reaction, Staining, Software, Co-Culture Assay, Flow Cytometry, MANN-WHITNEY

Journal: Nature Aging

Article Title: Preexisting senescent fibroblasts in the aged bladder create a tumor-permissive niche through CXCL12 secretion

doi: 10.1038/s43587-024-00704-1

Figure Lengend Snippet: a , Relative protein amounts of CXCL12 in transplanted bladder cancer from young (7–12-week-old) female p16-Cre ERT2 or p16-Cre ERT2 -DTR mice using cytokine array. (p16-Cre ERT2 , n = 3; p16-Cre ERT2 -DTR, n = 4). b , Representative IHC images of bladder with MB49 cells transplanted into young (7–12-week-old) female wild-type mice with vehicle (left top) or ABT-263 (left bottom) using an anti-MKI67 antibody. The percentage of MKI67 + cells is shown in the right panel. Scale bar, 100 μm. (n = 6 for each group). c , Representative IHC images of the bladders with MB49 cell transplantation in young (7–12-week-old) female p16-Cre ERT2 (top left) or p16-Cre ERT2 -DTR (bottom left) mice using an anti-CD105 antibody. The percentage of CD105 + area was shown in the right panel. Scale bar, 100 μm. (n = 6 for each group). d , Immunoblotting images using the indicated antibodies on the indicated cultured cell lines with or without treatment with CXCL12 recombinant protein. e , qPCR of Cxcr4 and Cxcr7 on the indicated cell lines. (n = 3 for each group). f , Experimental design of transplantation of the indicated cell lines and AMD3100 treatment using young (7–12-week-old) female wild-type mice. g , Immunoblotting images using lysates of bladder tumor originated from transplanted parental MB49 cells in young (7–12-week-old) female wild-type mice. Treatment with vehicle or AMD3100 was performed on the design of f . h , Experimental design of bladder allografts with the double treatment of tamoxifen+DT and AMD3100 using young (7–12-week-old) female p16-Cre ERT2 or p16-Cre ERT2 -DTR mice. i , Bladder weights on the design of h . (p16-Cre ERT2 , n = 6; p16-Cre ERT2 -DTR, n = 5). Unpaired t-test was performed for a–c and i . One-way ANOVA with Games-Howell’s test was performed for e . Data in a–c and e are presented as mean values ± SEM. For all box plots, notch of box blots indicates median value. Lower and upper bounds correspond to 25th and 75th percentiles, respectively. Cross mark indicates mean value. Whiskers extend to maximum or minimum value, with points outside this range drawn individually. All the t-test were two-side.

Article Snippet: Using the mouse cytokine antibody array kit panel A (#ARY006, R&D Systems), we evaluated cytokines in transplanted bladder cancer.

Techniques: Transplantation Assay, Western Blot, Cell Culture, Recombinant