Journal: Viruses

Article Title: Ephrin B1 and B2 Mediate Cedar Virus Entry into Egyptian Fruit Bat Cells

doi: 10.3390/v17040573

Figure Lengend Snippet: (q)PCR primers.

Article Snippet: Ephrin A2, A3, A4, B1 and B3 were synthesized at Eurofins Genomics, with the nucleotide sequences optimized according to human codon usage.

Techniques: Sequencing, Amplification

Journal: iScience

Article Title: TRPC4 deletion elicits behavioral defects in sociability by dysregulating expression of microRNA-138-2

doi: 10.1016/j.isci.2023.108617

Figure Lengend Snippet: Inhibition of miR-138 affects APT1 expression in Trpc4 −/− mice (A and B) Representative immunoblots and quantitative data of synaptic function-related proteins in the hippocampus. The level of APT1 was higher in the Trpc4 −/− hippocampus than in that of Trpc4 +/+ mice. However, the levels of PSD95, SYN1, and EphB3 were not significantly changed in Trpc4 −/− mice. (C) LNA-anti-miR-138 (anti-miR-138, 50 nM) was introduced into cultured hippocampal neurons and DIV13, and it reduced the endogenous level of miR-138 ( p < 0.001, n = 7). The expression of miR-138 was analyzed by qRT-PCR, and is expressed as fold changes relative to those in the anti-miR-CTL. (D and E) Representative immunoblots and quantitative data for APT1 normalized with the level of calnexin. Levels of APT1 are shown as fold changes relative to those of the anti-miR-CTL ( p < 0.01, n = 4). (F) The expression of Trpc4 mRNA measured by qRT-PCR was not changed by anti-miR-138. Expressions are indicated as fold changes relative to anti-miR-CTL. Each data point represents an individual sample; Data are means ± SEM. ∗∗ p < 0.01. ∗∗∗ p < 0.001. Unpaired t -tests. ns , not significant.

Article Snippet: Mouse anti-Ephrin B3 (EphB3) , Santa Cruz , Cat# sc271328; RRID: AB_10608991.

Techniques: Inhibition, Expressing, Western Blot, Cell Culture, Quantitative RT-PCR

Journal: iScience

Article Title: TRPC4 deletion elicits behavioral defects in sociability by dysregulating expression of microRNA-138-2

doi: 10.1016/j.isci.2023.108617

Figure Lengend Snippet: Behavioral phenotypes caused by Trpc4 deficiency are rescued by targeted overexpression of miR-138 in the hippocampal DG (A) Timeline of the experimental procedure. (B) Schematic illustration of control viral vectors (CTL) lenti-miR-138-2 (OE), and GFP immunostaining confirming localization of lentivirus infusion in the hippocampal DG. (C) Levels of miR-138 in the DG were upregulated in Trpc4 −/− , but not in Trpc4 +/+ , mice infused with lenti-miR-138-2 compared with lenti-CTL, respectively ( Trpc4 +/+ : p = 0.4693; Trpc4 −/− : p = 0.0008, n = 10–11 per group). (D) Representative immunoblots of APT1 and EphB3 in hippocampal DG lysates. (E) APT1 levels were reduced in mice expressing miR-138, while the levels of EphB3 were not significantly affected (APT1: p = 0.0023; EphB3: p = 0.125 n = 4 per group). (F) Grooming behavior. Lent-miR-138-2 did not significantly change grooming behavior in Trpc4 +/+ mice. However, it reduced time spent grooming in Trpc4 −/− mice ( ## p = 0.0022). (Genotype x Overexpression F 1,36 = 5.443, p = 0.0254, n = 10 per group). (G) Trpc4 +/+ mice infused with lenti-CTL and lenti-miR-138-2 displayed no significant difference in the rearing test ( Trpc4 +/+ mice, lenti-CTL vs. lenti-miR-138-2, p = 0.948). Lenti-miR-138-2 reduced time on rearing in Trpc4 −/− mice compared with those infused with lenti-CTL, recovered to levels in Trpc4 +/+ mice ( Trpc4 −/− , lenti-CTL vs . lenti-miR-138-2, p = 0.032; Genotype x overexpression F 1,36 = 2.65, p = 0.1123, n = 9–10 per group). (H) Social interaction test. Trpc4 +/+ mice infused with lenti-CTL and lenti-miR-138-2 showed a significant preference for S1 over the empty cage (lenti-CTL & lenti-miR-138-2, S1 vs . E, p < 0.0001, respectively), whereas Trpc4 −/− mice infused with lenti-CTL showed no preference for S1 relative to empty cage ( p = 0.9485). However, Trpc4 −/− mice, when infused with lenti-miR-138-2, spent more time sniffing with S1 than in an empty cage (Genotype x Overexpression F 3 , 80 = 2 . 8 , p = 0.0452, n = 10–13 per group). (I) Preference index. Trpc4 −/− mice infused with lenti-CTL showed less social interaction than Trpc4 +/+ controls ( ## p = 0.0009). However, when Trpc4 −/− mice were infused with lenti-miR-138-2 their preference index increased to the level of Trpc4 +/+ controls ( p = 0.0019). (Genotype x Overexpression F 1,40 = 5.928, p = 0.0195, n = 10–13 per group). (J) In the social novelty test, Trpc4 −/− mice infused with lenti-CTL displayed no significant preference for the S2 over the S1 ( p = 0.8896). However, when infused with lenti-miR-138-2 they had a preference for the S2 over the S1 ( p = 0.001). (Genotype × Overexpression interaction F 3 , 80 = 1.966, p = 0.1257, n = 10–13 per group). (K) The preference index for social novelty was significantly lower in Trpc4 −/− mice infused with lenti-CTL than in Trpc4 +/+ controls ( p = 0.0037), but it increased after lenti-miR-138-2 infusion in the Trpc4 −/− mice ( p = 0.0292). (Genotype × Overexpression interaction F 1,40 = 9.097, p = 0.0044, n = 10–13 per group). Each data point represents an individual mouse; Data are means ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; Unpaired t -tests for (C, E). Two-way ANOVA with Newman-Keuls multiple comparison post hoc test (F-K). ns , not significant. Source data were provided as a source data file. Scale bar: 50 μm.

Article Snippet: Mouse anti-Ephrin B3 (EphB3) , Santa Cruz , Cat# sc271328; RRID: AB_10608991.

Techniques: Over Expression, Control, Immunostaining, Western Blot, Expressing, Comparison

Journal: iScience

Article Title: TRPC4 deletion elicits behavioral defects in sociability by dysregulating expression of microRNA-138-2

doi: 10.1016/j.isci.2023.108617

Figure Lengend Snippet:

Article Snippet: Mouse anti-Ephrin B3 (EphB3) , Santa Cruz , Cat# sc271328; RRID: AB_10608991.

Techniques: Recombinant, Transfection, Lysis, Protease Inhibitor, Reverse Transcription, Control, Microarray, Gene Expression, Software

Journal: iScience

Article Title: TRPC4 deletion elicits behavioral defects in sociability by dysregulating expression of microRNA-138-2

doi: 10.1016/j.isci.2023.108617

Figure Lengend Snippet:

Article Snippet: Primary antibodies were diluted in 1X TBS with 0.1% (v/v) Tween-20, as follows: anti- APT1 (1:500, NBP2 17191, Novus), anti-EphB3 (1:1000, sc271328, Santa Cruz), anti-PSD95 (1:2000, #2507, Cell signaling), anti-Syn1 (1:2000, #5297, Cell signaling), anti-MATR3 (1:1000, ab70336, Abcam), anti-βactin (1:2000, sc47778, Santa Cruz), anti-GAPDH (1:1000, sc25778, Santa Cruz), anti- LaminB1 (1:1000, ab16048, Abcam).

Techniques: Recombinant, Transfection, Lysis, Protease Inhibitor, Reverse Transcription, Control, Microarray, Gene Expression, Software