Journal: Cell Death and Differentiation
Article Title: Neuronal MCT2 promotes angiogenesis via lactate in the developing mouse neocortex
doi: 10.1038/s41418-025-01581-w
Figure Lengend Snippet: a The schematic of the experimental protocol. Lentivirus of MCT2-shRNA (MCT2-shRNA) or scrambled shRNA particles, mixed with AAV-hSyn-GFP virus (to mark injected brain region), were injected into the S1 cortex of WT mice. b, c Representative immunostaining images of MCT1 + (cyan) BVs in the layers IV-V of viruses infected regions in the of S1 cortex in b , and quantification of MCT1 + -BV densities in layers IV-V ( n = 7 from 3 mice per group, mean ± SD, ** P < 0.01, one-way ANOVA with Tukey’s HSD test) in c were shown. d–f Representative images of co-immunostaining analysis of Podocalyxin (PODXL) and Isolectin GS-IB4 (red) BVs in layers IV-V of the S1 cortex in d , and quantifications of PODXL + and IB4 + BV densities in e , and PODXL + BV diameters in f (mean ± SD, n = 7 from 3 mice per group, ** P < 0.01, student’s t-test) were shown. g–i Representative images of co-immunostaining with indicated antibodies in g , and the quantifications (mean ± SD, n = 7 from 3 mice per group, ** P < 0.01, student’s t-test, or one-way ANOVA with Tukey’s HSD test) in h, i were shown. j, k Representative images of the co-immunostaining analysis using indicated antibodies in ( j ), and the quantifications of the phagocytic microglia (Iba1 + C-Casp-3 + ) densities (mean ± SD, n = 7 from 3 mice per group, ** P < 0.01, one-way ANOVA with Tukey’s HSD test) in k were shown. l, m Representative images of co-immunostaining analysis using indicated antibodies in l , and the quantifications of Egfl7 + cell density and ki67 + Egfl7 + over total ki67 + cells (%)(mean ± SD, n = 7 from 3 mice per group, * P < 0.05, ** P < 0.01, one-way ANOVA with Tukey’s HSD test) in m were shown. n A schematic of a working model. The blue dotted arrows indicate signaling pathways. Scale bars = 50 μm. The blue star (*) indicates a comparison between with WS or NS. The brown star (*) indicates a comparison between shRNA-MCT2 or scramble shRNA control, while the black star (*) indicates a comparison between each indicated experiment group, respectively.
Article Snippet: The following primary antibodies were used: rat monoclonal anti-CD31/PECAM (BD Biosciences, 553369); rabbit polyclonal anti-SLC16A1/MCT1 (Origene, TA321556); rabbit polyclonal anti- SLC16A7/MCT2 (Millipore Sigma, AB3542); rabbit polyclonal anti-SLC16A8/MCT3 (Novus Biologicals, NBP1-59885); mouse monoclonal anti-SMA-647 (Santa Cruz Biotechnology, sc-32251); chicken polyclonal anti-GFP (Aves Labs, GFP-1020); rat monoclonal anti-CD147/BSG (Bio-Red, MCA2283); isolectin GS-IB4, Alexa FluorTM 647 Conjugate (Invitrogen, 32450); guinea-pig polyclonal anti-NeuN (Millipore Sigma, ABN90); mouse monoclonal anti-β-actin (Cell Signal Technology, 3700S); rat monoclonal anti-mCherry (Invitrogen, M11217 ); rabbit monoclonal HIF1α (Cell Signal Technology, 3716S); rabbit monoclonal anti-HIF1α-OH (Cell Signal Technology, 3434S); rat monoclonal anti-Podocalyxin (PODXL, R&D system, 192703); rabbit polyclonal anti-Cleaved Casp-3 (Cell Signal Technology, 9661S); guinea-pig polyclonal anti-S100β (Synaptic systems, 287004); rabbit monoclonal anti-ERG-Alexa FluorTM 647 Conjugate (abcam, ab196149); rabbit polyclonal anti-Ki67(Millipore sigma, Ab9260); rat monoclonal anti-Ki67 (Synaptic Systems, HS-398 117); goat polyclonal anti-PDGFRβ (R&D Systems, AF1042); mouse anti-Glut1 (Millipore sigma, MABS132); mouse monoclonal anti-Oilg2 (EMD Millipore, MABN50); goat polyclonal anti-Iba1 (abcam, ab5076); Polyclonal Goat anti-Egfl7 (R&D Systems, AF3089); rabbit monoclonal anti-HIF2α (Cell Signal Technology, 71565S); rabbit monoclonal anti-VEGFR2 (Cell Signal Technology, 9698S); and rabbit monoclonal anti-pVEGFR2 (Cell Signal Technology, 2478S).
Techniques: shRNA, Virus, Injection, Immunostaining, Infection, Protein-Protein interactions, Comparison, Control