Journal: STAR Protocols

Article Title: Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing

doi: 10.1016/j.xpro.2025.104296

Figure Lengend Snippet: 3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Article Snippet: Collagen III antibody, anti-human, PE, REAdye_lease , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-357 RRID: AB_2905171.

Techniques: Imaging, Comparison, Selection, Staining

Journal: The Journals of Gerontology Series A: Biological Sciences and Medical Sciences

Article Title: Transcriptional Profiling of Muscle in Females With Distal Radius Fracture and Functional Sarcopenia

doi: 10.1093/gerona/glae002

Figure Lengend Snippet: Transcripts With Altered Gene Expression in the EXP Compared to NORM

Article Snippet: Recombinant human pro-collagen COL1A1 (R&D System, Minneapolis, MN, USA, 6220-CL) 75 μg/mL and recombinant human pro-collagen COL3A1 (R&D System, 7294-CL) 75 μg/mL were incubated with 5 μg/mL recombinant human BMP-1 (R&D, 1927-ZN) for 2 hours.

Techniques: Gene Expression

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Schematic representation of the human collagen XVIII variants, termed as SHORT, MIDDLE and LONG/FZ. Collagenous sequences are shown in white. Non-collagenous (NC) amino terminal sequences common to all variants are shown in black. Non-collagenous amino terminal sequences common to the two long variants are shown in grey. A non-collagenous amino terminal sequence specific to the LONG/FZ variant is marked with waves. The amino acid lengths (aa) and the predicted molecular masses (kDa) of these sequences are given, as are those of the full length type XVIII collagen molecules. Signal sequences are indicated by vertical and horizontal hatching. The epitopes of the antibodies used in this work are shown by black horizontal lines. The molecular masses of some endostatin-containing carboxy-terminal fragments are also indicated.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Sequencing, Variant Assay

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Immunoprecipitation with anti-ALL antibody and detection with anti-LONG huXVIII antibody. Elevated type XVIII collagen precursors in the plasma of ALI patients compared with normal controls is shown. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Immunoprecipitation, Clinical Proteomics

Journal: Critical Care

Article Title: Type XVIII collagen degradation products in acute lung injury

doi: 10.1186/cc7779

Figure Lengend Snippet: Western blot by anti-ALL huXVIII antibody. It shows elevated type XVIII collagen precursors in the bronchoalveolar lavage fluid (BALF) of patients with acute lung injury compared with normal controls. ALI = acute lung injury.

Article Snippet: A 25 μl sample of BALF was loaded onto polyacrylamide gel and the proteins were separated on a 7 to 12% SDS-PAGE under reducing conditions, electrotransferred to a nitrocellulose membrane (Protran; Schleicher & Schuell, Dassel, Germany) and probed with rabbit polyclonal antibodies against endostatin (HES.6) [ ] and human collagen XVIII (anti-all huXVIII QH4818 and anti-long huXVIII, QH1415.7) [ ], all used at a concentration 1 μg/ml in 5% fat-free milk powder in 1 × PBS, followed by a horseradish peroxidase-conjugated goat anti-rabbit antibody (Bio-Rad, Abingdon, UK).

Techniques: Western Blot

Journal: eLife

Article Title: Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

doi: 10.7554/elife.62927

Figure Lengend Snippet: Figure 6. NC410 increases collagen degradation products that correlate with tumor regression. (A) Schematic representation of the humanized murine tumor model used. HT-29 tumor was injected subcutaneously in the presence of human peripheral blood mononuclear cells. Mice were treated with NC410 or control by intraperitoneal injection, Q4D 4 doses followed by Q7D until endpoint. Mice were bled prior to start of experiment and weekly for 4 weeks. (B) Tumor growth kinetics with NC410. Asterisks indicate statistical significance: *p<0.05, ****p<0.0001, two-way ANOVA with Sidak multiple comparisons. (C) Analysis of collagen degradation products in serum at baseline, at weeks 1, 2, 3 and 4. reC1M: neo-epitope of MMP-2,9,13- mediated degradation of type I collagen; C3M: type III collagen degradation by MMP; C4M: type IV collagen degradation by MMP; C6M: neo-epitope of MMP-2-mediated degradation of type VI collagen; PRO-C3: pro-peptide of type III collagen/ECM formation/fibroblast activity; PRO-C6: pro-peptide of type VI collagen; VICM: neo-epitope of MMP-2,8, trypsin-mediated degradation of citrullinated vimentin; C4G: type IV collagen degraded by Figure 6 continued on next page

Article Snippet: The anti-human Fc antibody capture sensors (ForteBio) were first loaded with LAIR-2-Fc followed by an association step where the loaded sensor was dipped into wells containing human, mouse or rat collagen I (human, R&D Systems; mouse, Ray Biotech; rat, Yo Protein) or collagen III (human, R&D Systems; rat, Yo Protein).

Techniques: Injection, Control, Activity Assay