Journal: Bone Reports

Article Title: Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human induced pluripotent stem cells

doi: 10.1016/j.bonr.2024.101816

Figure Lengend Snippet: Three-dimensional osteogenic differentiation culture using human induced pluripotent stem cells (hiPSCs). ( a ) Schema showing three-dimensional osteogenic differentiation culture using hiPSCs. ( b ) Sections of the sponge were incubated for 28 days, and then stained with Alizarin Red S. Positive areas can be observed at the edges of the sponge. COL1A1 expression was analyzed using confocal laser microscopy. ( c ) Atelocollagen sponge without seeded hiPSCs. ( d)–(g ) Section of the sponge cultured for 28 days after seeding hiPSCs. Blue fluorescence indicates the nuclei and green fluorescence indicates COL1A1 expression. ( d ), ( e ) Cells stained only with DAPI without staining for COL1A1. ( e ) An enlarged image of a part of ( d ). Both ( d ) and ( e ) show the presence of nuclei at the edges and inside the sponge. ( f ), ( g ) Cells with additional staining for COL1A1. ( g ) An enlarged image of a region from ( f ). COL1A1-positive areas can be observed at the edges and inside the sponge. COL1A1, type I collagen alpha 1; hiPSC, human induced pluripotent stem cell.

Article Snippet: These sections were incubated with a primary antibody against type I collagen alpha 1 (COL1A1; diluted 1:100; 322-COLTt, PhosphoSolutions, Denver, CO, USA) at 37 °C in a humid chamber for 1 h. Subsequently, the sections were incubated at 25 °C for 1 h with the secondary antibody Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488; diluted 1:1000; Thermo Fisher Scientific).

Techniques: Incubation, Staining, Expressing, Microscopy, Cell Culture, Fluorescence

Journal: Bone Reports

Article Title: Identifying the best reference gene for RT-qPCR analyses of the three-dimensional osteogenic differentiation of human induced pluripotent stem cells

doi: 10.1016/j.bonr.2024.101816

Figure Lengend Snippet: Primers used for amplifying the target genes in the present study.

Article Snippet: These sections were incubated with a primary antibody against type I collagen alpha 1 (COL1A1; diluted 1:100; 322-COLTt, PhosphoSolutions, Denver, CO, USA) at 37 °C in a humid chamber for 1 h. Subsequently, the sections were incubated at 25 °C for 1 h with the secondary antibody Donkey Anti-Rabbit IgG H&L (Alexa Fluor® 488; diluted 1:1000; Thermo Fisher Scientific).

Techniques: Functional Assay, SYBR Green Assay, Marker, Gene Expression

Journal: Aging cell

Article Title: Unraveling Alveolar Fibroblast and Activated Myofibroblast Heterogeneity and Differentiation Trajectories During Lung Fibrosis Development and Resolution in Young and Old Mice.

doi: 10.1111/acel.14503

Figure Lengend Snippet: FIGURE 5 | Characterization of Ct1 and Ct2 cells in saline and Bleo-Tam d14 lungs. (a) RNA-scope of Cthrc1+ aMYF marker Cthrc1 and Ct1- cluster marker Inmt showing colocalization with tdTom+ cells. (b) RNA-scope of Cthrc1+ aMYF marker Cthrc1 combined with immunofluorescence (IF) for Pro-Col1a1 (Ct2-cluster marker), showing colocalization with tdTom+ cells. Scale bars: (a, b): 150 μm.

Article Snippet: Subsequently, the slides were then incubated overnight with anti- Pro- Col1a1 (Phosphosolutions, 321- COLP) and anti- tdTomato (Sicgen, AB8181- 200) at 4°C.

Techniques: Saline, RNAscope, Marker, Immunofluorescence