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. 2021 Oct 29;11:21307. doi: 10.1038/s41598-021-00232-7

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© The Author(s) 2021, corrected publication 2022

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Histological findings and RT-PCR gene expression results for the cartilage pellets on day 14 of co-culture. (a) HE staining for cartilage pellets on day 14 of co-culture. Left: Magnification: 40 × , Scale bars: 200 μm. Right: Magnification: 200 × , Scale bars: 100 μm. (b) TB staining for cartilage pellets on day 14 of co-culture. Left: Magnification: 40 × , Scale bars: 200 μm. Right: Magnification: 200 × , Scale bars: 100 μm. (c) Evaluation of the number of cartilage lacunae using TB staining. *P < 0.05, **P < 0.01, ***P < 0.0001. The data are shown as the mean ± SEM (n = 3). (d) Evaluation of the matrix proteins using immunohistochemical staining. Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. Magnification: 400 × , Scale bars: 100 μm. (e) Quantitative evaluation based on the DAB-positive cell count in immunohistochemical staining (on day 14 of co-culture). Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. **P < 0.01, ***P < 0.0001. NS: Not significant. The data are presented as the mean ± SEM (n = 3). (f) Gene expression analysis using RT-PCR (on day 14 of co-culture). Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. The data are presented as the mean ± SEM (n = 4).

Histological findings and RT-PCR gene expression results for the cartilage pellets on day 14 of co-culture. (a) HE staining for cartilage pellets on day 14 of co-culture. Left: Magnification: 40 × , Scale bars: 200 μm. Right: Magnification: 200 × , Scale bars: 100 μm. (b) TB staining for cartilage pellets on day 14 of co-culture. Left: Magnification: 40 × , Scale bars: 200 μm. Right: Magnification: 200 × , Scale bars: 100 μm. (c) Evaluation of the number of cartilage lacunae using TB staining. *P < 0.05, **P < 0.01, ***P < 0.0001. The data are shown as the mean ± SEM (n = 3). (d) Evaluation of the matrix proteins using immunohistochemical staining. Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. Magnification: 400 × , Scale bars: 100 μm. (e) Quantitative evaluation based on the DAB-positive cell count in immunohistochemical staining (on day 14 of co-culture). Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. **P < 0.01, ***P < 0.0001. NS: Not significant. The data are presented as the mean ± SEM (n = 3). (f) Gene expression analysis using RT-PCR (on day 14 of co-culture). Left: Collagen type 1, Middle: Collagen type 2, Right: Aggrecan. The data are presented as the mean ± SEM (n = 4).