integrin α6 cd49f  (Bio-Rad)

Journal: Nature aging

Article Title: Escape of hair follicle stem cells causes stem cell exhaustion during ageing

doi: 10.1038/s43587-021-00103-w

Figure Lengend Snippet: a, Images from a 5-hour movie of telogen HFs in control reveal no cell division and migration in the HF-SC compartment. Scale bar, 30μm. b, Images from a 6-hour movie of anagen HFs in control detect migrating HF-SCs and IRS cells. Red arrowhead indicates a downwardly migrating HF-SC and yellow arrowhead indicates an upwardly migrating IRS cell. Scale bar, 30μm. c, Images from a 4-hour movie of catagen HFs in dKO detect HF-SCs escaping from the bulge. Blue arrowhead indicates an HF-SC detaching from the bulge, red and green arrowheads indicate two HF-SCs squeezing through the basement membrane and escaping from the bulge region. Note the changed shape of nuclei during the escape. Scale bar, 10μm. d, Immunofluorescence signals of β4 integrin in dKO HFs. Arrowhead point to disrupted basement membrane with the loss of integrin staining. Scale bar, 20μm. e, Images from a 3.5-hour movie of a miniaturized hair follicle in dKO reveal the disintegration of HF-SCs, a cell division event and an escaped cell migrating in the dermis. Green arrowheads indicate two disintegrating cells in the miniaturized HF, red dashed circle indicates a dividing cell and yellow arrowhead indicates a migrating cell in the dermis. Scale bar, 20μm. f, Images from a 3.2-hour movie of a dying HF and a miniaturizing HF in dKO. Red arrowheads point to escaped epithelial cells with minimum activities in the dermis. Green arrowhead points to rapidly escaping cells from a miniaturizing HF. Scale bar, 50μm. g, A model illustrates HF-SC escape and HF miniaturization during ageing and in the dKO HF-SC compartment.

Article Snippet: The following antibodies were used: integrin α6 (CD49f, 1:75; eBioscience, PE-conjugated, 12–0495; APC-conjugated, 17–0495), CD34 (1:50; eBioscience, eFluor 660-conjugated, 50–0341), Sca1 (Ly-6A/E, 1:500; eBiosciences, PerCP-Cy5.5-conjugated, 45–5981).

Techniques: Control, Migration, Membrane, Immunofluorescence, Staining

Journal: Nature aging

Article Title: Escape of hair follicle stem cells causes stem cell exhaustion during ageing

doi: 10.1038/s43587-021-00103-w

Figure Lengend Snippet: a, Two-photon longitudinal tracking of hair follicles in young mice during the anagen to telogen hair cycle. Red numbers designate the same hair follicle in each image. Red dotted lines annotate the bulge region. Scale bar, 50μm. b, Two-photon intravital imaging of hair follicles from young (left panel) and old (middle and right panels) mice. White arrowheads point to miniaturized hair follicles and cells located outside of the HF-SC compartments. Red dotted lines outline miniaturized hair follicles. Scale bar, 50μm. c, Boxplot of the percentage of miniaturized hair follicles, quantified from 3-D scan of live animals. (n=205 HFs from 5 young mice; n=327 HFs from 3 old mice). d, Representative images of hair follicles with KRT5 and activated caspase 3 (acCas3) signals in young (6~8mo) and old (20mo) mice. (n=50 hair follicles from young mice; n=62 hair follicles from old mice, 3 pairs of mice). Scale bar, 20μm. e-f, Boxplot of number of acCas3+ HFSCs(e) and HG(f) per hair follicle (n=50 hair follicles from young mice; n=62 hair follicles from old mice, 3 pairs of mice). g, 3-D view of hair follicles in 24mo old mice. White arrowheads point to numerous escaped epithelial cells scattering in the dermis. Scale bar, 50μm. h, 3-D view of β4 integrin immunofluorescence signals in 24mo old mice. White arrowheads point to HF-SCs with protruding integrin signals in the new bulge side. Scale bar, 20μm.

Article Snippet: The following antibodies were used: integrin α6 (CD49f, 1:75; eBioscience, PE-conjugated, 12–0495; APC-conjugated, 17–0495), CD34 (1:50; eBioscience, eFluor 660-conjugated, 50–0341), Sca1 (Ly-6A/E, 1:500; eBiosciences, PerCP-Cy5.5-conjugated, 45–5981).

Techniques: Imaging, Immunofluorescence

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Spatial Engineering of Mammary Epithelial Cell Cultures with 3D Bioprinting Reveals Growth Control by Branch Point Proximity

doi: 10.1007/s10911-024-09557-1

Figure Lengend Snippet: Extrusion 3D bioprinting of mammary epithelial cells inside an ECM leads to formation of polarized 3D cell cultures of a defined shape. A-B Immunofluorescence imaging of the BM protein laminin α5 (LAMA5, green) and nuclei (blue) in clonal spheroid cultures of MCF10A cells A , and 3D-bioprinted cultures of MCF10A and MCF10DCIS.com cells B on day 14. Magnified images are shown in the ROIs. The MCF10A spheroids embedded in an ECM with elevated BME content A produced results comparable to the ECM used in B . C-D Immunofluorescence imaging of the basal epithelial integrin α6 (ITGA6, green) and nuclei (blue) in clonal spheroid cultures of MCF10A cells C , and 3D-bioprinted cultures of MCF10A and MCF10DCIS.com cells D on day 14. Magnified images are shown in the ROIs. The MCF10A spheroids embedded in an ECM with elevated BME content C produced results comparable to the ECM used in D . E Immunofluorescence imaging of E-cadherin (green), vimentin (magenta) and nuclei (blue) in 3D-bioprinted cultures of MCF10A and MCF10DCIS.com cells on day 10. Magnified images are shown in the ROIs. F Immunofluorescence imaging of nuclei (DAPI) in the 3D-bioprinted cultures of MCF10A and MCF10DCIS.com cells on day 14. Some of the apoptotic cells with fragmented nuclei are indicated with yellow arrowheads. Scale bars A, C: 100 μm, ROI 25 μm; B, D, E: 500 μm, ROI 50 μm; F: 50 μm. Images represent the central plane of the cell cultures

Article Snippet: The gels were incubated with primary antibodies against integrin α6 (NKI-GoH3, MCA699GA, Bio-Rad, 1:800), laminin α5 (4C7, ab17107-1001, Abcam, 1:50), E-cadherin (24E10, 3195 S, Cell Signaling, 1:200), vimentin (V9, 347 M-1, Sigma, 1:1000) and Ki67 (GR3375556-1, ab15580, Abcam, 1:250) in the blocking solution for 3 days at 4 °C on a shaker.

Techniques: Immunofluorescence, Imaging, Produced