Journal: Scientific Reports

Article Title: Collagenase-based wound debridement agent induces extracellular matrix supporting phenotype in macrophages

doi: 10.1038/s41598-024-53424-2

Figure Lengend Snippet: Volcano plot analysis of RT2 Profiler & RTPCR data reveals upregulation of collagen subtypes following CS-API treatment. ( a ) Schematic diagram of the validation of scRNA seq observation of Col1A1 expressing macrophage subtype using RT2 profiler and real time PCR analysis. ( b ) The volcano plot displays the differential gene expression analysis results of RT2 Profiler data, comparing CS-API-treated samples to control samples. Each data point represents a transcript, with the x-axis representing the log2 fold change and the y-axis representing the statistical significance (negative log10 p-value) of differential expression. The yellow data points indicate transcripts with a significant upregulation following CS-API treatment. Among the differentially expressed transcripts, COL1A1 stands out as one of the most abundantly upregulated transcripts. ( c ) Independent RT-PCR analysis was performed to validate the upregulation of COL1A1 mRNA following CS-API treatment. Expression levels of Col1A1 were measured using specific primers, and the results confirmed a significant increase in mRNA expression compared to control (TESCA buffer, vehicle) samples. Data presented as mean ± SEM (n = 6). * p < 0.05 compared to vehicle (control) treated MDM.

Article Snippet: Following fixation, cells were washed with PBS, blocked in 10% normal goat serum (NGS) for 30 min, and were incubated in primary antibodies Col1A1 (1:200; NBP1-77458F; FITC tagged; Novus), CD68 (1:200; ab213363, Abcam).

Techniques: Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Expressing, Real-time Polymerase Chain Reaction, Gene Expression, Control, Quantitative Proteomics

Journal: Scientific Reports

Article Title: Collagenase-based wound debridement agent induces extracellular matrix supporting phenotype in macrophages

doi: 10.1038/s41598-024-53424-2

Figure Lengend Snippet: Independent immunohistochemistry validation of Col1A1 protein expression. ( a-c ) The immunocytochemistry (IHC) analysis involved the detection of Col1A1 protein using specific antibodies (Col1A1, green and CD68, red, marker for mature macrophages) using fluorescence microscopy. ( a , b ) microscopy images of MDM cells treated with either ( a ), control buffer (vehicle, TESCA); or ( b ), CS-API; ( c ) The bar graph presents the quantification of the IHC images. Scale bar = 10 mm. au = arbitrary units. ( d ) Col1A1 protein expression was determined by enzyme-linked immunosorbent assay (ELISA). Data are mean ± SEM from 3–5 independent IHC images in each group. * p < 0.05.

Article Snippet: Following fixation, cells were washed with PBS, blocked in 10% normal goat serum (NGS) for 30 min, and were incubated in primary antibodies Col1A1 (1:200; NBP1-77458F; FITC tagged; Novus), CD68 (1:200; ab213363, Abcam).

Techniques: Immunohistochemistry, Biomarker Discovery, Expressing, Immunocytochemistry, Marker, Fluorescence, Microscopy, Control, Enzyme-linked Immunosorbent Assay