Fig 1: Immunocytostaining of neuronal (A) and glial (B) cell cultures of different brain regions: HT, HIP, OB, and RET. Anti-β-III tubulin (β-TUB III) was the primary antibody used to stain neurons, anti-GFAP and anti-AQP4 were used to label astrocytes, and anti-LSR Sigma was used to identify LSR protein. Images were taken using Fvi10 confocal microscope at a 40-X magnification (bar scale = 20 μm).
Fig 2: LSR is mainly expressed in glial cells in the CB.(A) Immunocytostaining of primary mixed, pure neuronal and glial cerebellar cell cultures using anti- β-III tubulin (β-TUB III) for neurons, anti-GFAP for astrocytes, and anti-LSR X-25 for LSR. Images were taken using Fvi10 confocal microscope at a 40-X magnification (bar scale = 20 μm). (B) Boxplot representation of expression ratio of total lsr, α, α’, and β in cerebellar glial and neuronal cell cultures relative to mixed cerebellar cultures. Statistical significance is represented as: * p ≤ 0.05, ** p≤0.01, *** p ≤ 0.001.
Fig 3: Working hypothesis of LSR effect on ApoE output in astrocytes. Our data show that high extracellular levels of lipoprotein (HM) stimulate lsr expression and decrease ApoE levels, suggesting a negative retro control of ApoE release from astrocytes, thus modulating ABCA1-mediated lipidation of lipoproteins. On the other hand, lsr reduction (siRNA) leads to release of lipidated ApoE and cholesterol into the medium. Transcriptional regulation of lsr by the LXR/RXR pathway depends on the lipoprotein content in the medium, while LXR upregulation of abca1 is not. We propose that the decoupling of cholesterol and ApoE release could influence the composition and lipidation of LP released by astrocytes. (LP = lipoprotein). Created with Biorender.com (accessed on 19 July 2022).
Fig 4: Age dependent changes in LSR protein levels in C57Bl/6JRj male mice.A, B, C, D, E, and F correspond to results obtained for hypothalamus (HT), hippocampus (HIP), olfactory bulb (OB), retina (RET), cortex (CX), and cerebellum (CB), respectively. In each panel there are immunoblots to detect LSR in the different brain structures, calculated percentages of α/α’ and β compared to total LSR, and expression ratio of total LSR with respect to β-TUB from young 3-month old (n = 3), and old 18-month old (n = 3). Statistical significance is represented as: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.
Fig 5: LSR and ABCA1 localization in astrocytes. LSR (A,B) and ABCA1 (C,D) protein levels were detected by immunofluorescence (red) in astrocytes grown in normal growth media (NM) in absence (A,C) or presence (B,D) of LXR agonist treatment (8 h, 1 µM T0901317). Co-labelling with DAPI to stain cell nuclei (blue) is shown in all panels (Scale bar = 10 µm). Arrows show strongly labelled cells; arrowheads show cells with discrete patchy or focal staining.
Supplier Page from MilliporeSigma for Anti-LSR antibody produced in rabbit