Journal: Cell Death & Disease

Article Title: Neuroprotective effects of bone marrow Sca-1 + cells against age-related retinal degeneration in OPTN E50K mice

doi: 10.1038/s41419-021-03851-0

Figure Lengend Snippet: A The mRNA levels of neurotrophic factors (NFs) were determined by real-time qPCR in both young and old wild-type (WT) and OPTN E50K mutant (E50K) mice. The mRNA expression of brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), fibroblast growth factor 2 (FGF2), and insulin-like growth factor 1(IGF-1) significantly decreased with age and was most dramatically decreased in EO retinas. WY: young WT; EY: young E50K. B – E The protein expression of BDNF, CNTF, FGF2, and IGF-1 in the retinas of the four groups was determined by ELISA. The protein levels of these NFs significantly decreased with age and were most dramatically decreased in the EO group ( n = 4–6/group). F – J Retinal sections were immunostained for BDNF, CNTF, FGF2, and IGF-1 in all 4 groups. The cell nuclei were visualized with DAPI. The intensity of staining of these NFs was significantly lower in the EO group than in the other 3 groups. Scale bar: 50 µm ( n = 3/group). The data are shown as the mean ± SEM. * P < 0.05, # P < 0.01.

Article Snippet: The membranes were then blocked with 5% skim milk solution for 1 h at room temperature before incubation with primary antibodies against BDNF (Proteintech, 28205), CNTF (Santa Cruz Biotechnology, 365210), FGF2 (Santa Cruz Biotechnology, 136255), IGF-1 (Abcam, 9572), Bcl-2 (Proteintech, 26593), and Bax (Proteintech, 50599) overnight at 4 °C.

Techniques: Mutagenesis, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Cell Death & Disease

Article Title: Neuroprotective effects of bone marrow Sca-1 + cells against age-related retinal degeneration in OPTN E50K mice

doi: 10.1038/s41419-021-03851-0

Figure Lengend Snippet: A The mRNA levels of NFs were determined by real-time qPCR in the bone marrow (BM) of the four groups. The mRNA expression of BDNF, CNTF, FGF2, and IGF-1 significantly decreased with age and was most obviously decreased in EO mice. B – E The protein expression of BDNF, CNTF, FGF2, and IGF-1 in the BM of the four groups was determined by ELISA. The protein levels of these NFs were significantly lower in the EO group than in the other 3 groups. ( n = 5–6/group). The data are shown as the mean ± SEM. * P < 0.05, # P < 0.01.

Article Snippet: The membranes were then blocked with 5% skim milk solution for 1 h at room temperature before incubation with primary antibodies against BDNF (Proteintech, 28205), CNTF (Santa Cruz Biotechnology, 365210), FGF2 (Santa Cruz Biotechnology, 136255), IGF-1 (Abcam, 9572), Bcl-2 (Proteintech, 26593), and Bax (Proteintech, 50599) overnight at 4 °C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cell Death & Disease

Article Title: Neuroprotective effects of bone marrow Sca-1 + cells against age-related retinal degeneration in OPTN E50K mice

doi: 10.1038/s41419-021-03851-0

Figure Lengend Snippet: A Bone marrow (BM)-derived Sca-1 + and Sca-1 − cells were immunostained for Sca-1 to confirm the phenotype. B – E The mRNA expression of NFs was determined by qPCR. The mRNA levels of BDNF, FGF2, CNTF, and IGF-1 were dramatically higher in Sca-1 + cells from young WT mouse BM than in Sca-1 − and total BM cells or Sca-1 + cells from young E50K mouse BM. Representative Western blot images and quantification of BDNF ( F, G ), FGF2 ( H, I ), CNTF ( J, K ), and IGF-1 ( L, M ) from Sca-1 + , Sca-1 − and total cells from both young WT and E50K mouse bone marrow. The reagent only (Veh) was used for the control group. GAPDH was used as a loading control ( n = 3/group). The data are shown as the mean ± SEM. * P < 0.05 , # P < 0.01 .

Article Snippet: The membranes were then blocked with 5% skim milk solution for 1 h at room temperature before incubation with primary antibodies against BDNF (Proteintech, 28205), CNTF (Santa Cruz Biotechnology, 365210), FGF2 (Santa Cruz Biotechnology, 136255), IGF-1 (Abcam, 9572), Bcl-2 (Proteintech, 26593), and Bax (Proteintech, 50599) overnight at 4 °C.

Techniques: Derivative Assay, Expressing, Western Blot, Control

Journal: Cell Death & Disease

Article Title: Neuroprotective effects of bone marrow Sca-1 + cells against age-related retinal degeneration in OPTN E50K mice

doi: 10.1038/s41419-021-03851-0

Figure Lengend Snippet: Retinal explants from old E50K mice were cultured with young BM-derived Sca-1 + and Sca-1 - cells of WT mice, and untreated E50K retinal explants (E50K) acted as the control group. The mRNA and protein expression of BDNF, CNTF, FGF2, and IGF-1 was determined by qPCR ( A ) and Western blot analysis ( B-F ) in old E50K retinal explants cocultured with Sca-1 + and Sca-1 - cells. Representative Western blot images ( G ) and the relative amounts of Bcl-2 ( H ) and Bax ( I ) are shown. J , K Retinal sections were immunostained for TUNEL in the 3 groups. The cell nuclei were visualized with DAPI. The number of TUNEL + cells in the ganglion cell layer (GCL) was significantly decreased in the Sca-1 + cell group ( n = 3/group). The data are shown as the mean ± SEM. * P < 0.05 , # P < 0.01 .

Article Snippet: The membranes were then blocked with 5% skim milk solution for 1 h at room temperature before incubation with primary antibodies against BDNF (Proteintech, 28205), CNTF (Santa Cruz Biotechnology, 365210), FGF2 (Santa Cruz Biotechnology, 136255), IGF-1 (Abcam, 9572), Bcl-2 (Proteintech, 26593), and Bax (Proteintech, 50599) overnight at 4 °C.

Techniques: Cell Culture, Derivative Assay, Control, Expressing, Western Blot, TUNEL Assay

Journal: Cell Death & Disease

Article Title: Neuroprotective effects of bone marrow Sca-1 + cells against age-related retinal degeneration in OPTN E50K mice

doi: 10.1038/s41419-021-03851-0

Figure Lengend Snippet: BM-derived Sca-1 + or Sca-1 − cells from young GFP (green fluorescent protein, green) transgenic mice were used to reconstitute old E50K mice, generating Sca-1 + and Sca-1 - chimaeras, respectively. A , B Retinal sections of Sca-1 + and Sca-1 − chimaeras were immunostained for GFP to confirm the capability of BM-derived Sca-1 + or Sca-1 − cells to home to retinas. C , D Retinal sections were immunostained with TUNEL (red) and for NeuN (green) in the 3 groups. The cell nuclei were visualized with DAPI. The number of TUNEL + cells in the GCL was significantly decreased in Sca-1 + chimaeras ( n = 3/group). E – H The protein levels of BDNF, CNTF, FGF2, and IGF-1 in the retinas of Sca-1 + and Sca-1 − chimaeras and age-matched E50K mice were determined by ELISA ( n = 3/group). The data are shown as the mean ± SEM. * P < 0.05, # P < 0.01.

Article Snippet: The membranes were then blocked with 5% skim milk solution for 1 h at room temperature before incubation with primary antibodies against BDNF (Proteintech, 28205), CNTF (Santa Cruz Biotechnology, 365210), FGF2 (Santa Cruz Biotechnology, 136255), IGF-1 (Abcam, 9572), Bcl-2 (Proteintech, 26593), and Bax (Proteintech, 50599) overnight at 4 °C.

Techniques: Derivative Assay, Transgenic Assay, TUNEL Assay, Enzyme-linked Immunosorbent Assay