Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: Ctdsp2 increases neural progenitor (NP) proliferation. (A–C) Overexpression of Ctdsp2 , but not the control construct pCAGIG, increased the proportion of cells expressing both proliferative marker bromodeoxyuridine + (BrdU + )/green fluorescence protein + (GFP + ) and radial glial cell (RGC) marker Pax6 + /GFP + , but not intermediate progenitor (IP) marker Tbr2 + /GFP + , in GFP-positive cells. (D–F) shRNA-mediated knockdown (sh Ctdsp2 ) of Ctdsp2 decreased the proportion of both BrdU + /GFP + cells and Pax6 + /GFP + cells, but not Tbr2 + /GFP + cells in GFP-positive cells, compared to the control construct pSilencer. Values represent mean ± SEM. n = 9 sections from at least three brains. ** P < 0.01; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, Fluorescence, shRNA, Knockdown

Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: miR-26 promotes NP proliferation. (A,D,G) Overexpression of miR-26a, but not the control construct pCAGIG, proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + , but not IP marker Tbr2 + /GFP + , in GFP-positive cells. (B,E,H) miRNA sponge-mediated knockdown (miR-26-SP), but not the mutated sponge (miR-26-SPmut), decreased proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + in GFP-positive cells. (C,F,I) Ratio of BrdU + /GFP + , Pax6 + /GFP + or Tbr2 + /GFP + cells vs. GFP + cells in the electroporated cortex. Values represent mean ± SEM. n = 9 sections from at least three brains. * P < 0.05; ** P < 0.01; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, Knockdown

Journal: Frontiers in Molecular Neuroscience

Article Title: Cohesive Regulation of Neural Progenitor Development by microRNA miR-26, Its Host Gene Ctdsp and Target Gene Emx2 in the Mouse Embryonic Cerebral Cortex

doi: 10.3389/fnmol.2018.00044

Figure Lengend Snippet: Emx2 is functionally inhibited by miR-26 in regulating NP proliferation. (A–F) Overexpression of Emx2 , but not the control construct pCAGIG, decreased the proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + , but not IP marker Tbr2 + /GFP + , in GFP-positive cells. short hairpin RNA (shRNA)-mediated knockdown (sh Emx2 ) of Emx2 increased the proportion of both BrdU + /GFP + cells and Pax6 + /GFP + cells, but not Tbr2 + /GFP + cells in GFP-positive cells, compared to the control construct pSilencer. (G–J) Emx2 expression suppressed the proportion of cells expressing both proliferative marker BrdU + /GFP + and RGC marker Pax6 + /GFP + in GFP-positive cells. Co-expressing Emx2 with miR-26, but not miR-26-mut, dramatically reversed the suppression. (K,L) Emx2 expression did not alter the proportion of cells expressing IP marker Tbr2 + /GFP + , in GFP-positive cells. Values represent mean ± SEM. n = 9 sections from at least three brains. ** P < 0.01; *** P < 0.001; ns, not significant. ANOVA with post hoc test was used. Scale bar = 50 μm.

Article Snippet: Primary antibodies against the following antigens were used: BrdU (1:50, Developmental Studies Hybridoma Bank at University of Iowa (DSHB)), Ki67 (1:500, Abcam), Pax6 (1:500, Covance), Pax6 (1:15 DSHB), Tbr2 (1:500, Abcam), GFP (1:1000, Abcam, chicken), and GFP (1:1000, Rockland, rabbit).

Techniques: Over Expression, Control, Construct, Expressing, Marker, shRNA, Knockdown

Journal: Neurotherapeutics

Article Title: Down-regulation of lipocalin-2 alleviates depressive-like behaviors in mice through modulation of microglial activation

doi: 10.1016/j.neurot.2026.e00862

Figure Lengend Snippet: LCN2 neutralizing antibodies alleviated the depressive-like behaviors in CUS mice. ( A ) Schematic of the experimental procedure and behavioral studies. ( B ) Levels of LCN2 measured by ELISA in the mouse hippocampus treated with IgG or anti-LCN2 (n = 6/group). ∗∗∗ P < 0.001 versus the IgG group using Student's t -test. ( C – H ) LCN2 neutralizing antibodies relieved depressive-like behaviors in CUS mice (n = 12/group) as measured by the SPT ( C ), FST ( D) , TST ( E ) and OFT ( F – H ). ∗∗∗ P < 0.001 versus the Control + IgG group; ### P < 0.001 versus the CUS + anti-IgG group using two-way ANOVA followed by the Holm-Sidak post hoc multiple comparison test.

Article Snippet: After incubation in blocking buffer, membranes were incubated overnight at 4 °C with the following primary antibodies: anti-LCN2 (1:1000, abs134038, Absin), anti-iNOS (1:1000, 18985-1-AP, Proteintech Group), anti-p(Ser616)DRP1 (1:1000, ab314755, Abcam), anti-DRP1 (1:1000, 12957-1-AP, Proteintech Group), anti-MFN2(1:1000, 12186-1-AP, Proteintech Group), and anti-GAPDH (1:3000, 60004-1-AP, Proteintech Group).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison