Journal: Journal of Biomedical Research
Article Title: HDGF derived from Müller cells enhances the activation of microglia in diabetic retinopathy
doi: 10.7555/JBR.38.20240386
Figure Lengend Snippet: High glucose upregulated the expression of HDGF receptor in microglia. A: Western blotting analysis of HDGF protein expression in BV2 microglial cells cultured under low-glucose (LG, 1 g/L) or high-glucose (HG, 5 g/L) conditions for 48 h. α-Tubulin served as the loading control. Quantitative protein levels of ITGB2 in BV2 and Müller cells ( n = 3 per group). B: Representative immunofluorescence images of retinal cryosections from control and streptozotocin (STZ)-induced diabetic mice, co-stained for HDGF (red) and the microglial marker IBA1 (green). Nuclei were counterstained with DAPI (blue). Assessment of immunofluorescence colocalization of HDGF and IBA1 in the two experimental groups (left, control; right, STZ). C: Single-cell RNA sequencing analysis of control and STZ-diabetic mouse retinas. UMAP plot showing the expression of Ncl (the gene encoding nucleolin) across all cells (upper). The microglia cluster is indicated. Dot plot illustrating the relative expression level and proportion of Ncl -expressing cells in microglia in the control and STZ groups (lower). D: Representative immunofluorescence images (left) and quantification of mean fluorescence intensity (MFI, right) for NCL (green) in BV2 cells cultured under LG or HG conditions for 48 h. Nuclei were counterstained with DAPI (blue). n = 3. Data are presented as mean ± standard deviation. Statistical significance was determined by the paired Student's t -test. * P < 0.05. Abbreviations: HDGF, hepatoma-derived growth factor; IBA1, ionized calcium-binding adaptor molecule 1; ns, not significant; STZ, streptozotocin.
Article Snippet: The immortalized murine microglial cell line (BV2) (Cat. #CRL-2469) was purchased from the American Type Culture Collection and cultured in DMEM/F12 medium (Gibco) supplemented with 10% FBS (Invitrogen), 1% glutamine (Invitrogen), and streptomycin (100 U/mL; Invitrogen) at 37 °C, 5% CO 2 .
Techniques: Expressing, Western Blot, Cell Culture, Control, Immunofluorescence, Staining, Marker, RNA Sequencing, Fluorescence, Standard Deviation, Derivative Assay, Binding Assay