anti ck18 antibody  (Bioss)

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Representative trajectories of Cy5-proKPV NPs and Cy5-labeled PS NPs in the mouse intestinal mucus, as determined by MTrackJ analysis of 5-min confocal microscopy recordings. ( B ) Time-dependent ensemble-averaged geometric MSD of proKPV NPs and PS NPs in the mouse intestinal mucus. ( C ) Distributions of log( D eff ) for proKPV NPs and PS NPs at a 5-min timescale, with at least 100 particles tracked per sample for each type. ( D ) Three-dimensional fluorescence images illustrate the penetration of Cy5-KPV and Cy5-proKPV NPs through the mouse intestinal mucus. ( E ) Representative ex vivo images (left) and quantitative analysis (right) illustrating accumulation of Cy5-labeled proKPV in the colons of mice with or without DSS-induced acute colitis at 6 hours after oral administration. ( F and G ) Fluorescence images (F) and quantitative analysis (G) of the colonic tissues isolated from colitis mice at different time points following treatment with free Cy5-KPV or Cy5-proKPV. ( H ) AUC of Cy5 fluorescence in colonic tissues after treatment with free Cy5-KPV or Cy5-proKPV. ( I ) Fluorescence images of cryosections of colonic tissues from colitis mice at 6 hours after treatment with Cy5-KPV or Cy5-proKPV. Nuclei were stained with DAPI (blue). ( J to L ) Immunofluorescence analysis of colocalization of Cy5-KPV or Cy5-proKPV with CK18 + intestinal epithelial cells (J), CD68 + macrophages (K), or Ly6G + neutrophils (L) in cryosections of colonic tissues. Data in (E), (G), and (H) are presented as means ± SD ( n = 3 biological replicates). * P < 0.05; *** P < 0.001.

Article Snippet: Also, colonic tissue cryosections were prepared and stained with anti-CK18 antibody (bsm-52058R, Bioss Biotechnology, China), anti-occludin antibody (27260-1-AP, Proteintech Biotechnology, China), anti–ZO-1 antibody (21773-1-AP, Proteintech Biotechnology, China), anti-Ly6G antibody (562737, BD Biosciences, USA), anti-CD206 antibody (ab64693, Abcam, USA), anti-CitH3 antibody (ab281584, Abcam, USA), anti-NE antibody (ab314916, Abcam, USA), or anti-FOXP3 antibody (ab215206, Abcam, USA).

Techniques: Labeling, Confocal Microscopy, Fluorescence, Ex Vivo, Isolation, Staining, Immunofluorescence

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Schematic illustration of treatment regimens. ( B ) Body weight of mice during a 7-day treatment course. Data were normalized as the percentages of the body weight at day 0.** P < 0.01; *** P < 0.001; n.s., not significant versus the colitis group. ( C ) Changes in DAI. ** P < 0.01 and *** P < 0.001 versus the colitis group. ( D ) Digital photos (left) and quantified lengths (right) of colonic tissues isolated from mice at day 7 following different treatments. Scale bars, 5 mm. ( E ) Representative miniendoscopic images of colons from mice after 7 days of treatment. ( F ) Colonic tissue sections stained with H&E or PAS for different groups. ( G ) Immunofluorescence analysis of the expression of CK18, occludin, and ZO-1 in colonic tissues from mice following various treatments. Scale bars, 100 μm. ( H to M ) Levels of TNF-α (H), IL-1β (I), IL-6 (J), MPO (K), MDA (L), and ROS (M) in colonic tissues isolated from healthy or diseased mice treated with different formulations. After 7 days of treatment, colonic tissue homogenates were prepared, and mediator concentrations were quantified and normalized to the total protein content. ( N ) Immunofluorescence of colonic tissues showing Ly6G + neutrophils, CD206 + M2 macrophages, CitH3/NE-positive NETs, and FOXP3 + T reg cells after different treatments. Data in (B) to (D) and (H) to (M) are presented as means ± SD ( n = 6 biological replicates). * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant.

Article Snippet: Also, colonic tissue cryosections were prepared and stained with anti-CK18 antibody (bsm-52058R, Bioss Biotechnology, China), anti-occludin antibody (27260-1-AP, Proteintech Biotechnology, China), anti–ZO-1 antibody (21773-1-AP, Proteintech Biotechnology, China), anti-Ly6G antibody (562737, BD Biosciences, USA), anti-CD206 antibody (ab64693, Abcam, USA), anti-CitH3 antibody (ab281584, Abcam, USA), anti-NE antibody (ab314916, Abcam, USA), or anti-FOXP3 antibody (ab215206, Abcam, USA).

Techniques: Isolation, Staining, Immunofluorescence, Expressing

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Body weight changes of mice during 7-day treatment, which were normalized to the baseline at day 0. ** P < 0.01 and *** P < 0.001; n.s., not significant versus the colitis group. ( B ) Changes in DAI values. ** P < 0.01 and *** P < 0.001 versus the colitis group. ( C and D ) Digital photos (C) and quantified lengths (D) of colonic tissues isolated from mice after 7 days of treatment. ( E ) Histological sections of colonic tissues stained with H&E or PAS. ( F ) Immunofluorescence indicates expression patterns of CK18, occludin, and ZO-1 in colonic tissues. ( G to J ) Levels of TNF-α (G), IL-6 (H), IL-1β (I), and MPO (J) in colonic tissues isolated from healthy or diseased mice treated with different formulations. ( K and L ) Immunofluorescence analysis of NETs (K), Ly6G + neutrophils, and CD206 + macrophages (L) in colonic tissues. Data in (A), (B), (D), and (G) to (J) are presented as means ± SD ( n = 6 biological replicates). * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant.

Article Snippet: Also, colonic tissue cryosections were prepared and stained with anti-CK18 antibody (bsm-52058R, Bioss Biotechnology, China), anti-occludin antibody (27260-1-AP, Proteintech Biotechnology, China), anti–ZO-1 antibody (21773-1-AP, Proteintech Biotechnology, China), anti-Ly6G antibody (562737, BD Biosciences, USA), anti-CD206 antibody (ab64693, Abcam, USA), anti-CitH3 antibody (ab281584, Abcam, USA), anti-NE antibody (ab314916, Abcam, USA), or anti-FOXP3 antibody (ab215206, Abcam, USA).

Techniques: Isolation, Staining, Immunofluorescence, Expressing

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Representative trajectories of Cy5-proKPV NPs and Cy5-labeled PS NPs in the mouse intestinal mucus, as determined by MTrackJ analysis of 5-min confocal microscopy recordings. ( B ) Time-dependent ensemble-averaged geometric MSD of proKPV NPs and PS NPs in the mouse intestinal mucus. ( C ) Distributions of log( D eff ) for proKPV NPs and PS NPs at a 5-min timescale, with at least 100 particles tracked per sample for each type. ( D ) Three-dimensional fluorescence images illustrate the penetration of Cy5-KPV and Cy5-proKPV NPs through the mouse intestinal mucus. ( E ) Representative ex vivo images (left) and quantitative analysis (right) illustrating accumulation of Cy5-labeled proKPV in the colons of mice with or without DSS-induced acute colitis at 6 hours after oral administration. ( F and G ) Fluorescence images (F) and quantitative analysis (G) of the colonic tissues isolated from colitis mice at different time points following treatment with free Cy5-KPV or Cy5-proKPV. ( H ) AUC of Cy5 fluorescence in colonic tissues after treatment with free Cy5-KPV or Cy5-proKPV. ( I ) Fluorescence images of cryosections of colonic tissues from colitis mice at 6 hours after treatment with Cy5-KPV or Cy5-proKPV. Nuclei were stained with DAPI (blue). ( J to L ) Immunofluorescence analysis of colocalization of Cy5-KPV or Cy5-proKPV with CK18 + intestinal epithelial cells (J), CD68 + macrophages (K), or Ly6G + neutrophils (L) in cryosections of colonic tissues. Data in (E), (G), and (H) are presented as means ± SD ( n = 3 biological replicates). * P < 0.05; *** P < 0.001.

Article Snippet: The obtained cryosections were separately stained with anti–cytokeratin 18 (CK18) antibody (bsm-52058R, Bioss Biotechnology, China), anti-F4/80 antibody (30325, Cell Signaling Technology, USA), or anti-Ly6G antibody (31469, Cell Signaling Technology, USA) to further evaluate the tissue distribution of NPs.

Techniques: Labeling, Confocal Microscopy, Fluorescence, Ex Vivo, Isolation, Staining, Immunofluorescence

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Schematic illustration of treatment regimens. ( B ) Body weight of mice during a 7-day treatment course. Data were normalized as the percentages of the body weight at day 0.** P < 0.01; *** P < 0.001; n.s., not significant versus the colitis group. ( C ) Changes in DAI. ** P < 0.01 and *** P < 0.001 versus the colitis group. ( D ) Digital photos (left) and quantified lengths (right) of colonic tissues isolated from mice at day 7 following different treatments. Scale bars, 5 mm. ( E ) Representative miniendoscopic images of colons from mice after 7 days of treatment. ( F ) Colonic tissue sections stained with H&E or PAS for different groups. ( G ) Immunofluorescence analysis of the expression of CK18, occludin, and ZO-1 in colonic tissues from mice following various treatments. Scale bars, 100 μm. ( H to M ) Levels of TNF-α (H), IL-1β (I), IL-6 (J), MPO (K), MDA (L), and ROS (M) in colonic tissues isolated from healthy or diseased mice treated with different formulations. After 7 days of treatment, colonic tissue homogenates were prepared, and mediator concentrations were quantified and normalized to the total protein content. ( N ) Immunofluorescence of colonic tissues showing Ly6G + neutrophils, CD206 + M2 macrophages, CitH3/NE-positive NETs, and FOXP3 + T reg cells after different treatments. Data in (B) to (D) and (H) to (M) are presented as means ± SD ( n = 6 biological replicates). * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant.

Article Snippet: The obtained cryosections were separately stained with anti–cytokeratin 18 (CK18) antibody (bsm-52058R, Bioss Biotechnology, China), anti-F4/80 antibody (30325, Cell Signaling Technology, USA), or anti-Ly6G antibody (31469, Cell Signaling Technology, USA) to further evaluate the tissue distribution of NPs.

Techniques: Isolation, Staining, Immunofluorescence, Expressing

Journal: Science Advances

Article Title: Inflammation-triggered self-immolative conjugates enable oral peptide delivery by overcoming gastrointestinal barriers

doi: 10.1126/sciadv.aea2989

Figure Lengend Snippet: ( A ) Body weight changes of mice during 7-day treatment, which were normalized to the baseline at day 0. ** P < 0.01 and *** P < 0.001; n.s., not significant versus the colitis group. ( B ) Changes in DAI values. ** P < 0.01 and *** P < 0.001 versus the colitis group. ( C and D ) Digital photos (C) and quantified lengths (D) of colonic tissues isolated from mice after 7 days of treatment. ( E ) Histological sections of colonic tissues stained with H&E or PAS. ( F ) Immunofluorescence indicates expression patterns of CK18, occludin, and ZO-1 in colonic tissues. ( G to J ) Levels of TNF-α (G), IL-6 (H), IL-1β (I), and MPO (J) in colonic tissues isolated from healthy or diseased mice treated with different formulations. ( K and L ) Immunofluorescence analysis of NETs (K), Ly6G + neutrophils, and CD206 + macrophages (L) in colonic tissues. Data in (A), (B), (D), and (G) to (J) are presented as means ± SD ( n = 6 biological replicates). * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant.

Article Snippet: The obtained cryosections were separately stained with anti–cytokeratin 18 (CK18) antibody (bsm-52058R, Bioss Biotechnology, China), anti-F4/80 antibody (30325, Cell Signaling Technology, USA), or anti-Ly6G antibody (31469, Cell Signaling Technology, USA) to further evaluate the tissue distribution of NPs.

Techniques: Isolation, Staining, Immunofluorescence, Expressing