rat cd146 apc  (Miltenyi Biotec)

Journal: Journal of Advanced Research

Article Title: Chaperone-mediated autophagy sustains pericyte stemness necessary for brain tissue homeostasis

doi: 10.1016/j.jare.2025.04.015

Figure Lengend Snippet: Donor PC therapy restores CMA in host PCs in damaged tissue. (A) Representative images showing co-localization of puncta pattern expression of LAMP-2A protein (green) with a PC marker, α-SMA (red), analyzed in host PCs of microvessels (v) of control mice, in mice brain with lesioned areas (injured) and treated intravenously with control adMSCs and with donor brain WT or KO PCs. Nuclei were stained with DAPI (blue). Scale bar: 100 µm. Right panels show bigger magnification of double positive cells. ( B ) Representative images showing co-localization of puncta pattern expression of LAMP-2A protein (in green) with a PC marker, PDGFRβ (in red), analyzed in host PCs of microvessels (v) of control mice, in mice brain with lesioned areas and treated intravenously with control adMSCs and with donor brain WT or KO PCs. Right panels show bigger magnification of double positive cells. Nuclei were stained with DAPI (blue). Scale bars: 100 µm. (C) Representative images showing co-localization of puncta pattern expression of LAMP-2A protein (in green) with a PC marker, CD146 (in red), analyzed in host PCs of microvessels (v) of control mice, in mice brain with lesioned areas and treated intravenously with donor brain PC. Right panels show bigger magnification of double positive cells (Scale bars: 20 µm). Nuclei were stained with DAPI (blue). Scale bars: 100 µm. ( D ) Relative quantification of percentage of PCs in brain microvessels expressing the PC marker α -SMA (n = 40 α -SMA + PCs per condition), PDGFRβ (n = 20 PDGFRβ + PCs per condition), or CD146 (n = 35 CD146 + PCs per condition) and the CMA receptor LAMP-2A (PC/LAMP-2A +/+ ) over total PCs. All data represent mean ± SD obtained from at least, three experiments represented as dots, independently; * p < 0.05; ** p < 0.01; *** p < 0.001; ns indicates no statistical significance (ANOVA with Tukey’s post-test). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For fluorescent double labelling, mouse anti-alpha-smooth muscle actin (α-SMA; Abcam, ab7817), goat anti-platelet derived growth factor receptor beta (PDGFRβ; R&D Systems, BAF1042), or rat CD146-APC (Miltenyi Biotec, 130–118-408) antibodies were used in combination with rabbit anti-LAMP-2A (Invitrogen, 51–2200) antibody.

Techniques: Expressing, Marker, Control, Staining, Quantitative Proteomics

Journal: International Journal of Molecular Sciences

Article Title: Endothelial Colony-Forming Cells Dysfunctions Are Associated with Arterial Hypertension in a Rat Model of Intrauterine Growth Restriction

doi: 10.3390/ijms221810159

Figure Lengend Snippet: ECFC quantification. Flow cytometry analysis of cultured cells was performed on CTRL-ECFCs and IUGR-ECFCs isolated from six-month-old male rats. ( A – D ). Left panel; FSC versus SSC plot. Cells were gated to exclude subcellular debris. Right panel; CD45 versus DAPI staining on gated cells from left panel ( A ). Dead cells (DAPI+) and hematopoietic cells (CD45+) were excluded by gating CD31+ versus CD146+ staining on CD45-viable cells ( B ). Upper panel, cells from IUGR rats, lower-left three panels from CTRL rats, and values were reported in the histogram ( D ). Negative control stain on CD45-viable cells (in the absence of CD31 and CD146) was represented in the lower-right panel ( C ); * p < 0.05.

Article Snippet: Single-cell suspensions from CTRL-ECFCs ( n = 3) and IUGR-ECFCs ( n = 5) were stained with fluorochrome-labeled monoclonal antibodies against CD31 PE (TLD-3A12), CD45 FITC (OX-1), and CD146 (LSEC) APC (BD Biosciences, San Jose, CA, USA; or Miltenyi Biotech, Bergisch Gladbach, Germany) in PBS/3%FCS for 20 min at 4 °C.

Techniques: Flow Cytometry, Cell Culture, Isolation, Staining, Negative Control