Biased development of granulocytic MDSCs in Rip2-deficient mice.

<p>A, Inflammatory cytokine expression in serum of Rip2^+/+ and Rip2^−/− mice intravesically implanted with MB49 cells for 12 days was assessed by cytokine array analysis. Cytokines increased in Rip2^−/− mice are depicted in the schematic as shown. (+) 2-5x, (++) 5-10x, (+++) >10x based on quantitation by ImageJ. Results are from duplicates and representative of two independent experiments. B, Splenocytes from Rip2^+/+ and Rip2^−/− mice were examined for expression of CD4, CD8, B220, NK1.1, while CD11b⁺ cells were examined for expression of Ly6G and Ly6C by flow cytometry as indicated. Column, mean of four mice; bars SD. Data are representative of two independent experiments. C, Total tumor from Rip2^+/+ and Rip2^−/− mice intravesically implanted with MB49 cells and sacrificed at 12 days were examined for expression of IL4, IFNγ, M-CSF, G-CSF, and GM-CSF by qPCR. Column, mean of three mice; bars SD. D, Tumors from Rip2^+/+ and Rip2^−/− mice subcutaneously implanted with MB49 cells for 14 days were fractioned to CD45⁻ and CD45⁺CD11b⁺Gr1^hi cells and examined for G-CSF expression by qPCR. Column, mean of three mice; bars SD. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Loss of Rip2 alters composition of tumor infiltrating cells from Rip2-deficient mice.

<p>Infiltration of CD8, CD4, NK, CD11b, F4/80, Ly6G, and Ly6C expressing cells as indicated, were examined by immunofluorescence, and Foxp3 by immunohistochemistry in bladder tumors from Rip2^+/+ and Rip2^−/− mice intravesically implanted with MB49 cells and sacrificed at 12 days. Left panels show specific antibody staining, middle panel shows DAPI staining, right panel shows merging of antibody and DAPI stains. Representative examples from each group of four to six mice in 4 independent experiments are shown at x40 magnification. Bar graphs enumerate mean number of cells per x40 field of 3 representative sections from groups of four mice; bars, SD. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Intrinsic enhanced development of MDSC in absence of Rip2.

<p>A, Bone marrow from naïve Rip2^+/+ and Rip2^−/− mice were differentiated by M-CSF, G-CSF, GM-CSF, and G-CSF plus GM-CSF as indicated for four days prior to flow cytometry to examine expression of Ly6G and Ly6C in CD11b⁺ cells (ranging from 70–90%, not shown). CD11b⁺LyG⁺Ly6C^lo and CD11b⁺LyG^loLy6C^hi populations for each group are shown as total cell counts per 10⁵ starting BM cells. B, Expression of arginase-1 and iNOS by qPCR in G-CSF and G-CSF plus GM-CSF differentiated bone marrow from naïve Rip2^+/+ and Rip2^−/− mice. Column, mean of three mice; bars SD. Data are representative of two independent experiments. Representative examples are shown. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Increased tumor infiltrating granulocytic MDSCs in Rip2-deficient mice.

<p>Rip2^+/+ and Rip2^−/− mice subcutaneously implanted with MB49 cells for 14 days were assessed for (A) tumor weight, and sorted by flow cytometry to examine (B) CD45⁺NK1.1⁺ cells, and (C) CD45⁺CD11b⁺Gr1^hi and CD45⁺CD11b⁺Gr1^lo cells. Column, mean of four mice; bars SD. Data are representative of two independent experiments. D, Sorted CD45⁺CD11b⁺Gr1^hi and CD45⁺CD11b⁺Gr1^lo cells in Rip2^+/+ and Rip2^−/− mice were examined for expression of arginase-1 and iNOS by qPCR, while CD45⁺CD11b⁺Gr1^hi cells in Rip2^+/+ and Rip2^−/− mice were examined for expression of IL10, IL-12, and IFNγ by qPCR. Column, mean of two mice; bars SD. Data are representative of three independent experiments. Representative examples are shown. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Increased tumor size and metastases in Rip2-deficient mice.

<p>Rip2^+/+ and Rip2^−/− mice intravesically implanted with MB49 cells and sacrificed at 12 days were assessed for bladder tumor (A) weight, (B) histology with H&E staining at x5 and x20 magnification. C, Lungs were examined for metastasis by number of metastatic lesions per coronal cross section, % incidence, histology with H&E staining at x10 and x40 magnification, and gross examination. D, Kidneys were examined for metastasis by number of metastatic lesions per coronal cross section, % incidence, and histology with H&E staining at x5 and x20 magnification. Representative examples are shown. Columns, mean of 33 Rip2^+/+ and 19 Rip2^−/− mice pooled from 5 independent experiments; bars, SEM. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Enhanced epithelial-to-mesenchymal transition in tumors from Rip2-deficient mice.

<p>A, Rip2^+/+ and Rip2^−/− mice intravesically implanted with MB49 cells for 12 days were assessed for E-cadherin and N-cadherin expression by immunofluorescence. Magnification of x40. Data are representative of two independent experiments. B, Bar graphs below enumerate mean number of cells per x40 field of 3 representative sections from groups of four mice; bars, SD. C, Expression of <i>zeb-1</i>, <i>zeb-2</i>, <i>snail</i>, and <i>twist</i> were examined from CD45⁻ dissociated total tumor tissue by qPCR. Column, mean of four mice; bars SD. Data are representative of two independent experiments. D, Expression of <i>zeb-1</i>, <i>zeb-2</i>, <i>snail</i>, and <i>twist</i> were examined from dissociated lungs containing metastases by qPCR. Column, mean of four mice; bars SD. All <i>p</i> values were determined by two-tailed Student's <i>t</i> test, with statistically significance defined as p<0.05.</p

Unlocking Chemically Encrypted Information Using Three Types of External Stimuli

Encryption is critical to information security; however, existing chemical-based information encryption strategies are still in their infancy. We report here a new approach to chemical encryption involving a supramolecular gel QR (quick response) code with multiple encryption functions. Three color “turn-on” supramolecular polymer gels, G1–G3, were prepared that produce pink, purple, and yellow colors when subject to treatment with acetic acid vapor, UV light, and methanolic FeCl3, respectively. As the result of hydrogen-bonding interactions at the gel interfaces, the three gels can be assembled to produce gel G4. Engraving a QR code pattern onto G4 then gave gel G5. When one or two stimuli are applied to the individual pieces corresponding to the QR engraved versions of the gels G1–G3 making up G5, a complete scannable pattern is not displayed, and the stored information cannot be recognized. Only when three different stimuli are applied at the same time does G5 give a complete recognizable pattern allowing the stored information to be retrieved. This strategy was applied to the decryption-based opening of a coded lock

Unlocking Chemically Encrypted Information Using Three Types of External Stimuli

Encryption is critical to information security; however, existing chemical-based information encryption strategies are still in their infancy. We report here a new approach to chemical encryption involving a supramolecular gel QR (quick response) code with multiple encryption functions. Three color “turn-on” supramolecular polymer gels, G1–G3, were prepared that produce pink, purple, and yellow colors when subject to treatment with acetic acid vapor, UV light, and methanolic FeCl3, respectively. As the result of hydrogen-bonding interactions at the gel interfaces, the three gels can be assembled to produce gel G4. Engraving a QR code pattern onto G4 then gave gel G5. When one or two stimuli are applied to the individual pieces corresponding to the QR engraved versions of the gels G1–G3 making up G5, a complete scannable pattern is not displayed, and the stored information cannot be recognized. Only when three different stimuli are applied at the same time does G5 give a complete recognizable pattern allowing the stored information to be retrieved. This strategy was applied to the decryption-based opening of a coded lock

Unlocking Chemically Encrypted Information Using Three Types of External Stimuli

Encryption is critical to information security; however, existing chemical-based information encryption strategies are still in their infancy. We report here a new approach to chemical encryption involving a supramolecular gel QR (quick response) code with multiple encryption functions. Three color “turn-on” supramolecular polymer gels, G1–G3, were prepared that produce pink, purple, and yellow colors when subject to treatment with acetic acid vapor, UV light, and methanolic FeCl3, respectively. As the result of hydrogen-bonding interactions at the gel interfaces, the three gels can be assembled to produce gel G4. Engraving a QR code pattern onto G4 then gave gel G5. When one or two stimuli are applied to the individual pieces corresponding to the QR engraved versions of the gels G1–G3 making up G5, a complete scannable pattern is not displayed, and the stored information cannot be recognized. Only when three different stimuli are applied at the same time does G5 give a complete recognizable pattern allowing the stored information to be retrieved. This strategy was applied to the decryption-based opening of a coded lock

Unlocking Chemically Encrypted Information Using Three Types of External Stimuli

Encryption is critical to information security; however, existing chemical-based information encryption strategies are still in their infancy. We report here a new approach to chemical encryption involving a supramolecular gel QR (quick response) code with multiple encryption functions. Three color “turn-on” supramolecular polymer gels, G1–G3, were prepared that produce pink, purple, and yellow colors when subject to treatment with acetic acid vapor, UV light, and methanolic FeCl3, respectively. As the result of hydrogen-bonding interactions at the gel interfaces, the three gels can be assembled to produce gel G4. Engraving a QR code pattern onto G4 then gave gel G5. When one or two stimuli are applied to the individual pieces corresponding to the QR engraved versions of the gels G1–G3 making up G5, a complete scannable pattern is not displayed, and the stored information cannot be recognized. Only when three different stimuli are applied at the same time does G5 give a complete recognizable pattern allowing the stored information to be retrieved. This strategy was applied to the decryption-based opening of a coded lock