mouse balb cj strain 000651  (Jackson Laboratory)

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: Identification of IP6K3 as a responsive gene in MASLD and exercise training. A Volcano plot showing differentially expressed genes in the bariatric intervention cohort ( GSE48452 ); B volcano plot showing differentially expressed genes in the NASH cohort ( GSE89632 ); C scatter plot illustrating oppositely regulated genes between bariatric intervention and NASH datasets; D intersection analysis of genes upregulated or downregulated in bariatric intervention and NASH cohorts; E , G KEGG pathway enrichment analysis of overlapping genes in bariatric intervention ( E ) and NASH ( G ) datasets; F , H gene set enrichment analysis showing enrichment of inositol phosphate metabolism and phosphatidylinositol signaling system pathways; I immunoblot and qPCR analysis of IP6K3 expression in liver samples from patients with or without MASLD; J immunoblot and qPCR analysis of IP6K3 expression in livers from normal diet (ND) and HFHC-fed mice; K analysis of GSE189600 (publicly available single-nucleus RNA-sequencing data from the livers of patients with MASLD or healthy individuals) revealed that IP6K3 was abundantly expressed in hepatic parenchymal cells; L analysis of odds ratios (OR) and a plot of IP6K3 transcriptional levels in various cell types in GSE189600 . The red arrow indicates hepatocytes; M immunofluorescent staining of IP6K3 (green) with Alb (red, a marker of hepatocytes) and CD68 (red, a marker of macrophages in human) in liver tissues of patients with MASLD. Immunofluorescent staining of IP6K3 (green) with Alb (red) and F4/80 (red, a marker of macrophages in mice) in the liver tissues of mice fed an HFHC diet. DAPI was used to stain nuclei (blue). Scale bar, 100 μm; N schematic illustration of the HFHC diet-induced MASLD model with or without exercise training; O immunoblot and qPCR analysis of IP6K3 expression in liver samples from HFHC mice with or without exercise training; P immunofluorescent staining of IP6K3 (green) with Actin (red) in the liver tissues from HFHC mice with or without exercise training. DAPI was used to stain nuclei (blue). scale bar, 100 μm. n = 4–6 per group, ** P < 0.01, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Analysis, Western Blot, Expressing, RNA Sequencing, Staining, Marker

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: Hepatocyte-specific Ip6k3 deletion attenuates HFHC-induced liver injury. Four groups of Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice fed a normal diet (ND) or an HFHC diet were included, and the following indices were examined. A Representative images and quantitative analyses of HE, Masson’s trichrome, Oil Red O, and periodic acid–Schiff (PAS) staining in the liver tissues. Scale bar, 100 µm; B , C GTT and ITT assay; D serum ALT, AST, TG, and TC levels; E mRNA levels of pro‐inflammatory factors ( Il-6 , Il-1β , Tnf-α , Mcp-1 ), lipid synthesis ( Acca , Fas , Srebp1c ), lipid intake ( Fabp1 , Fatp1 , Cd36 , Ppar-γ ), bile acid synthesis ( Cyp7a1 ), and fatty acid oxidation ( Ucp2 , Pdk4 , Ppara , Cpt1a , Acox1 ) in the liver tissues; F protein levels of α-SMA, TNF-α, IL-6, and IL-1β in the liver tissues; G protein levels of p-AKT and p-IRS1 in the liver tissues. n = 3–8 per group, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Staining

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: Single-cell RNA sequencing reveals that hepatocyte-specific Ip6k3 deletion reprograms ferroptosis and BCAA metabolism. Single-cell RNA sequencing (scRNA-seq) analysis was performed with liver tissues from Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice fed an HFHC diet. A Schematic illustration of the experimental workflow for scRNA-seq; B UMAP visualization of ten cell clusters; C heatmap showing the expression of the indicated genes in ten different cell clusters; D relative proportions of the ten major liver cell populations in HFHC-fed Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice; E intercellular interactions among ten cell clusters; F volcano plot showing the downregulated (197) and upregulated (240) genes in Ip6k3 fl / fl Alb-Cre mice fed an HFHC diet when compared with the Ip6k3 fl / fl counterpart; G KEGG enrichment analysis showing the upregulated (red) and downregulated (blue) genes in hepatocytes in Ip6k3 fl / fl Alb-Cre mice fed an HFHC diet when compared with the Ip6k3 fl / fl counterpart. H UMAP visualization of hepatocyte subsets; I Trajectory inference of hepatocytes along the predominant transcriptional trajectory (partition 1) under HFHC conditions; J Pseudotime-dependent changes in iron-, ferroptosis resistance- and BCAA degradation-associated gene signature scores along the predominant hepatocyte trajectory (partition 1) in HFHC-fed Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice. Gene signature scores were calculated based on curated gene sets associated with iron handling, ferroptosis regulation, and BCAA degradation

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Single Cell, RNA Sequencing, Analysis, Expressing

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: Hepatocyte-specific Ip6k3 deletion and exercise training suppress ferroptosis and accelerate BCAA metabolism in MASLD. A Four groups were designed: Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice fed a normal diet (ND) or an HFHC diet. mRNA levels of pro-ferroptotic ( Tfr, Acsl4, Alox15 ) and anti-ferroptotic factors ( Gpx4, Dhodh, Fsp1 ) in liver tissues of the four groups; B protein levels of pro-ferroptotic (TFR, ACSL4, ALOX15) and anti-ferroptotic factors (GPX4, DHODH, FSP1) in liver tissues of the four groups; C liver GSH, the ratio of NAD + /NADH and activity of SOD in liver tissues of the four groups; D , E ROS production and free iron in living cells (mostly Fe 2+ ) assessed by 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) or FerroOrange probes with flow cytometry in single‐cell suspensions prepared from mice liver tissues of the four groups; F , G serum BCAA levels measured by LC–MS, and liver BCAA levels assessed by commercial BCAA assay kit in the four groups; H three groups were designed: Ip6k3 fl / fl mice without exercise training, Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice with 8-week exercise training. All three groups were fed a 16-week HFHC diet; I liver GSH, NAD + /NADH, and activity of SOD in three groups; J , K mRNA levels of pro-ferroptotic ( Tfr , Acsl4 , Alox15 ) and anti-ferroptotic factors ( Gpx4 ), and corresponding protein levels in liver tissues of the three groups; L , M Serum (detected by LC–MS) and liver (detected by a BCAA assay kit) BCAA levels in the three groups. n = 3–10 per group, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Activity Assay, Flow Cytometry, Single Cell, Liquid Chromatography with Mass Spectroscopy

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: IP6K3 negatively regulated BCAT2 expression in MASLD. A and D mRNA levels of Bcat1 , Bcat2 , Ppm1k , Bckdk , Bckdhα , and Bckdhβ and protein level of BCAT2 in liver tissues of Ip6k3 fl / fl and Ip6k3 fl / fl Alb-Cre mice fed a normal diet (ND) or an HFHC diet; B and E mRNA levels of BCAT1 , BCAT2 , PPM1K , BCKDK , BCKDHα , and BCKDHβ and protein level of BCAT2 in HepG2 cells treated with si-scramble or si- IP6K3 with or without PA incubation; C and F mRNA levels of Bcat1 , Bcat2, Ppm1k , Bckdk , Bckdhα , and Bckdhβ and protein level of BCAT2 in liver tissues of Ip6k3 fl / fl , Ip6k3 fl / fl + exercise and Ip6k3 fl / fl Alb-Cre + exercise mice fed with HFHC diet. n = 3 for protein, n = 6 for mRNA, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Expressing, Incubation

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: IP6K3 regulates BCAA-induced ferroptosis in a BCAT2-dependent manner. A – C Representative images and quantification of lipid peroxidation assessed by C11-BODIPY (581/591) staining, mRNA levels of pro-ferroptotic ( TFR , ACSL4 , ALOX15 ) and anti-ferroptotic factors ( GPX4 ) and the corresponding protein levels in HepG2 cells transfected with pcDNA3.1 without BCAA stimulation, and pcDNA3.1 or pcDNA3.1- IP6K3 with BCAA stimulation; D – F Representative images and quantification of lipid peroxidation assessed by C11-BODIPY (581/591) staining, mRNA levels of pro-ferroptotic ( TFR , ACSL4 , ALOX15 ) and anti-ferroptotic factors ( GPX4 ) and the corresponding protein levels in HepG2 cells transfected with pcDNA3.1, or pcDNA3.1- IP6K3 with or without Fer-1 treatment (all three groups was under BCAA stimulation). G – I Representative images and quantification of lipid peroxidation assessed by C11-BODIPY (581/591) staining, mRNA levels of pro-ferroptotic ( TFR , ACSL4, , ALOX15 ) and anti-ferroptotic factors ( GPX4 ) and the corresponding protein levels in HepG2 cells transfected with si-scramble without BCAA stimulation, and si-scramble or si -IP6K3 with BCAA stimulation; J – L Representative images and quantification of lipid peroxidation assessed by C11-BODIPY (581/591) staining, mRNA levels of pro-ferroptotic ( TFR , ACSL4 , ALOX15 ) and anti-ferroptotic factors ( GPX4 ) and the corresponding protein levels in HepG2 cells transfected with si- scramble with or without iBCAT2 (BCAT-IN-2, a BCAT2 inhibitor) treatment, or transfected with si -IP6K3 under BCAA stimulation with or without iBCAT2 treatment. n = 3 for protein, n = 6 for mRNA and C11-BODIPY (581/591) staining. Scale bar, 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Staining, Transfection

Journal: Cellular & Molecular Biology Letters

Article Title: Exercise suppresses IP6K3 to modulate BCAA metabolism and ferroptosis in MASLD

doi: 10.1186/s11658-026-00981-1

Figure Lengend Snippet: IP6K3 interacts with HNRNPK to destabilize BCAT2 mRNA. A HepG2 cells expressing HA-tagged- IP6K3 were treated with PA, lysed, immunoprecipitated with anti-HA, and separated via SDS-PAGE. The most prominent ~ 65 kDa band (indicated by red box) was excised and subjected to mass spectrometry. Molecules interacting with IP6K3 were identified using a nano liquid chromatography–tandem mass spectrometry, and HNRNPK was selected for further investigation; B AlphaFold3-predicted interaction pattern between IP6K3 and HNRNPK. Binding affinity estimation using PRODIGY predicted a strong interaction between IP6K3 and HNRNPK (Δ G ≈ −23.1 kcal/mol); C Co-IP and immunoblotting detected the interaction between IP6K3 and HNRNPK in HepG2 cells under PA stimulation; D actinomycin D chase assay for BCAT2 mRNA expression in HepG2 cells transfected with either pcDNA3.1 or pcDNA3.1- HNRNPK with or without PA stimulation; E , F mRNA levels of BCAT2 and the corresponding protein level of BCAT2 in HepG2 cells treated with either pcDNA3.1 or pcDNA3.1- HNRNPK with or without PA stimulation; G A working model of IP6K3-regulated ferroptosis in MASLD. n = 3 for protein, n = 6 for mRNA, * P < 0.05, *** P < 0.001, **** P < 0.0001

Article Snippet: An Ip6k3 flox/flox mouse strain was generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9 technology: two pairs of sgRNA sequences targeting exon 3 of the Ip6k3 gene were designed using the CRISPR/Cas9 system; Cas9 mRNA and gRNA were synthesized through in vitro transcription and microinjected into fertilized eggs of C57BL/6J mice to generate F0 mice ( Ip6k3 flox/flox ).

Techniques: Expressing, Immunoprecipitation, SDS Page, Mass Spectrometry, Liquid Chromatography, Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Transfection

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: In vitro assessment of new AntagoNAT sequences (A) Schematic of AAV-CMV-eGFP-miR-155-AntagoNAT-miR-155-WPRE plasmid. (B) Transient transfection revealed 13 sequences to significantly increase Scn1a expression compared to the control cells (∗ p < 0.001). Candidates K, H, and J showed the highest increase in expression (16-fold compared to control). One-way ANOVA with multiple comparison correction using Two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli. n = 9/10 for AntagoNAT sequences, n = 6 for control (3 technical replicates each). (C) Schematic of AAV-U6-AntagoNAT-CMV-eGFP-WPRE plasmid. (D) In vitro transduction of AAV9 vectors ( n = 5/6 replicates of each group, 3 technical replicates each) using either the U6 or the CMV promoter, represented in a Boxplot. One-way ANOVA, Dunnett’s multiple comparison performed.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: In Vitro, Plasmid Preparation, Transfection, Expressing, Control, Comparison, Transduction

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Neonatal AAV9-AntagoNAT-H gene therapy increases survival and reduces febrile seizures in DS mice Scn1a +/− mice received either AAV9-AntagoNAT-H or PBS via i.c.v. and i.v. PBS was also administered to wild-type mice ( Scn1a +/+ ). (A) Experiment timeline. (B) Survival of Scn1a +/− DS mice treated with AAV9-AntagoNAT-H ( p = 0.04), shown in percentage of survival. Log-rank (Mantel-Cox) test. (C) Weight curves, data presented as mean, with SD error bars. Two-way ANOVA with Dunnett’s multiple comparisons test. (D) Febrile seizure temperature threshold assessment, showing a significant reduction with AAV9-AntagoNAT-H treated group ( p < 0.001). Log-rank (Mantel-Cox) test. (E) Duration of febrile seizures reduced in AAV9-AntagoNAT-H group ( p < 0.001). Man-Whitney test. (F) The Racine score classification of febrile seizures observed in the experimental mice. A significant reduction in seizure severity was observed in AAV9-AntagoNAT-H group ( p = 0.02). Fisher's exact test.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Increase of endogenous Scn1a in the cortex of AAV9-AntagoNAT-H-treated Scn1a +/− DS mice (A) Scn1a expression in the cortex, significantly increased with AAV9-AntagoNAT-H ( p = 0.032) and Scn1a +/+ PBS ( p = 0.003) compared to PBS group. (B) Scn1a expression in the heart showed a signifcant difference between Scn1a +/+ PBS and Scn1a +/− group ( p < 0.001) only. (C) Vector copy number analysis in the cortex showed a significant increase in AAV9-AntagoNAT-H group ( p < 0.001). (D) Vector copy number in the heart only revelaed a signifcant difference between Scn1a +/+ and Scn1a +/ − PBS groups ( p < 0.001). (E) Na V 1.1 expression in the cortex. One-way ANOVA Holm-Šídák’s multiple comparisons test. For each group, n values are indicated in the figure.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: Expressing, Plasmid Preparation

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV9-mediated targeting of natural antisense transcript as a novel treatment for Dravet syndrome

doi: 10.1016/j.omtn.2026.102942

Figure Lengend Snippet: Increase in endogenous Scn1a after i.c.v. and i.v. AAV9-AntagoNAT-H therapy to P14 Scn1a +/− DS mice (A) Schematic diagram showing the experimental plan. (B) Survival curve for AAV9-H dose D, showing percentage of survival. (C) Representative images of GFAP (top) and CD68 (middle) staining in the brain and CD68 in the liver (bottom) of animals that received AAV9-H dose D. Further details in . (D) Survival curve of animals that received AAV9-H dose E, showing percentage of survival. Log-rank (Mantel-Cox) test. (E) Scn1a expression in cerebral cortex displayed a significant increase in both Scn1a +/+ PBS and Scn1a +/ − AAV9-AntagoNAT-H groups ( p = 0.009) compared to Scn1a +/ − PBS group. Cerebral cortex tissues for Scn1a +/− group, range from P20 to 100. (F) Na V 1.1 expression in the cerebral cortex, reported a difference between control groups only ( p = 0.013). One-way ANOVA Holm-Šídák’s multiple comparisons test.

Article Snippet: 129 S- Scn1a tm1Kea /Mmjax DS mouse strain, which contain an exon 1 deletion of Scn1a , resulting in haploinsufficiency, were obtained from the Jackson Laboratory (Jackson Laboratory, ME, USA).

Techniques: Staining, Expressing, Control

Journal: Molecular Therapy Advances

Article Title: Adeno-associated virus-induced neurotoxicity is prevented by CpG depletion

doi: 10.1016/j.omta.2026.201703

Figure Lengend Snippet: CpG depletion does not alter transgene expression but prevents AAV-induced dendritic loss (A) Immunoblot of S1 homogenates probed with antibodies against transgene product EGFP and loading control GAPDH following injection of 1 × 10 10 vg of unmodified AAV1-pCAG-FLEX-EGFP or CpG-depleted AAV1-pCAG-FLEX-EGFP into Slc17a7-IRES2-Cre-D mice. Each lane contains samples from a different animal. UT, untreated. (B) Densitometry shows that EGFP protein levels are indistinguishable in ipsilateral S1 of Slc17a7-IRES2-Cre-D mice injected with either CpG-depleted or unmodified virus. EGFP protein levels are normalized to GAPDH protein levels in the same lane and represented as a fold change from the AAV1-treated condition. (C) Representative cortical neurons from untreated wild-type mice or ipsilateral (injected) hemispheres of wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Scale bars, 100 μm. (D) Total dendritic length and (E) total Sholl intersections of cortical neurons from untreated wild-type mice and wild-type mice injected with 1 × 10 10 vg of either unmodified or CpG-depleted AAV1-pCAG-FLEX-EGFP. Values are represented as a ratio of injected (ipsi) normalized to uninjected (contra) hemispheres. (F–H) Sholl intersections plotted as a function of distance from the soma for cortical neurons from the same animals shown in (E). (D and E), n = 4 untreated control mice, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with unmodified AAV1-pCAG-FLEX-EGFP, 10–14 neurons per animal (average = 12 cells); n = 5 mice injected with CpG-depleted AAV1-pCAG-FLEX-EGFP, 12–15 neurons per animal (average = 13 cells). Data in (B and D–H) are represented as mean ± SD. ∗∗∗∗ p < .0001; ∗∗∗ p < .001; ∗∗ p < .01; ns, not significant. In (G), a single ∗ indicates p < 0.05, and vertically stacked ∗ indicate p < 0.01. Statistical analysis: in (B, F–H), two-tailed unpaired t test; in (D) and (E), one-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: Male C57BL/6J and Slc17a7-IRES2-Cre-D (Strain # 037512) mice were obtained from Jackson Laboratory (The Jackson Laboratory, Bar Harbor, ME).

Techniques: Expressing, Western Blot, Control, Injection, Virus, Two Tailed Test, Comparison