Journal: Frontiers in Neuroscience

Article Title: Differentiation timing-dependent axon targeting and subtype specification in retinal ganglion cells

doi: 10.3389/fnins.2026.1733811

Figure Lengend Snippet: Axon projections of the neurogenic-tagged RGCs in the optic tract and LGN. (A) Schematic gene structures of the neurogenic tagging driver Neurod1 CreER (D1B) and R26-CAG-LF-mTFP1 reporter. Tamoxifen (TM) injection was administered at E11.5, E13.5, or E15.5 to induce CreER recombination. AAV2-CAG-FLPo was injected intravitreally at the postnatal stage to induce Flp recombination in an eye-specific manner. (B,C) Labeling of TM-tagged axon arbors by mTFP1 immunostaining (white in the upper panels, green in the lower panels) in brain sections. The TM-tagging stages are indicated at the top. DAPI staining is shown in blue. (B) Labeled axons in the contralateral optic tract (arrowheads) at P28 for TM11.5, P29 for TM13.5, and P32 for TM15.5. The dotted line indicates the optic tract border. (C) Labeled axons in the dorsal lateral geniculate nucleus (dLGN; arrowheads) and ventral LGN (vLGN; arrows) at P28 for TM11.5, P29 for TM13.5, and P32 for TM15.5. Contra, contralateral projections; Ipsi, ipsilateral projections. Scale bars, 250 μm.

Article Snippet: One to two microliters of AAV2-CAG-FLPo solution (4.5 × 10 12 genome copies/ml; Vector Biolabs, Cat# VB1313) was injected through the hole using a pulled and beveled glass pipette (Drummond Cat#5–000-1001-X10).

Techniques: Injection, Labeling, Immunostaining, Staining

Journal: Frontiers in Neuroscience

Article Title: Differentiation timing-dependent axon targeting and subtype specification in retinal ganglion cells

doi: 10.3389/fnins.2026.1733811

Figure Lengend Snippet: Axonal projections of the neurogenic-tagged RGCs in the MTN and SCN. (A,B) Labeling of TM-tagged axon arbors by mTFP1 staining (white in upper panels, green in lower panels) in brain sections using the same strategy as in : Neurod1 CreER (D1B) mice were crossed with R26-CAG-LF-mTFP1 reporter mice, followed by intravitreal injection of AAV2-CAG-FLPo into one eye at the postnatal stage. TM-tagging stages are indicated at the top. DAPI staining is shown in blue. (A) Labeled axons in the contralateral medial terminal nucleus (MTN), indicated by arrowheads, at P29 for TM11.5, P35 for TM13.5, and P32 for TM15.5. MTNd, dorsal MTN; MTNv, ventral MTN. (B) Suprachiasmatic nucleus (SCN) at P28 for TM11.5, P29 for TM13.5, and P32 for TM15.5. Right, contralateral side; left, ipsilateral side. The dotted line indicates the border of the SCN. Scale bars: 50 μm (A) and 250 μm (B) .

Article Snippet: One to two microliters of AAV2-CAG-FLPo solution (4.5 × 10 12 genome copies/ml; Vector Biolabs, Cat# VB1313) was injected through the hole using a pulled and beveled glass pipette (Drummond Cat#5–000-1001-X10).

Techniques: Labeling, Staining, Injection

Journal: Frontiers in Neuroscience

Article Title: Differentiation timing-dependent axon targeting and subtype specification in retinal ganglion cells

doi: 10.3389/fnins.2026.1733811

Figure Lengend Snippet: Axonal projections of the neurogenic-tagged RGCs in the SC. (A,B) TM-tagged axon arbors labeled by mTFP1 staining (white in left panels, green in right panels) in the superior colliculus (SC) using the same strategy as in : Neurod1 CreER (D1B) mice were crossed with R26-CAG-LF-mTFP1 reporter mice, followed by intravitreal injection of AAV2-CAG-FLPo into one eye at the postnatal stage. TM-tagging stages are indicated at the top. Brain sections were analyzed at P28 for TM11.5, P35 for TM13.5, and P32 for TM15.5. DAPI staining is shown in blue. Arrowheads indicate labeled axons in the ipsilateral SC. Arrows indicate labeled axons in the contralateral SC. Right, contralateral side; left, ipsilateral side. (A) Rostral SC, (B) Caudal SC. Scale bars, 500 μm.

Article Snippet: One to two microliters of AAV2-CAG-FLPo solution (4.5 × 10 12 genome copies/ml; Vector Biolabs, Cat# VB1313) was injected through the hole using a pulled and beveled glass pipette (Drummond Cat#5–000-1001-X10).

Techniques: Labeling, Staining, Injection

Journal: Scientific Reports

Article Title: LIMK2 inactivation suppresses mechanical stimulation-induced dermal fibroblast differentiation and resistance to apoptosis

doi: 10.1038/s41598-026-37610-y

Figure Lengend Snippet: ( a ) AAV2 expressing LIMK2-inactive (IA)-mutant-GFP and AAV2 expressing GFP (control) were used to infect normal HDFs. Expression levels of LIMK2 and GFP were analyzed by Western blotting. The original blots are presented in Supplementary Fig. 7. ( b ) Decreased baseline cofilin phosphorylation 48 h after LIMK2-IA-mutant-GFP infection. In HDFs and GFP-HDFs (both controls), mechanical stimulation for 30 min increased p-cofilin levels, whereas in LIMK2-IA-mutant-GFP-HDFs, p-cofilin levels remained unchanged. ( c ) In LIMK2-IA-mutant-GFP-HDFs, α-SMA expression was not changed at 24 h after mechanical stimulation. In contrast, in HDFs and GFP-HDFs, α-SMA expression was significantly increased by mechanical stimulation. ( d ) LIMK2 activation increased, whereas LIMK2 inactivation decreased, cofilin phosphorylation in HDFs and KDFs. ( e ) LIMK2 activation increased α-SMA expression in both HDFs and KDFs, whereas LIMK2 inactivation decreased α-SMA expression only in KDFs. ( f ) LIMK2 activation increased, whereas LIMK2 inactivation decreased, collagen type I expression in HDFs and KDFs. ** p < 0.01. * p < 0.05. ## p < 0.01 vs HDFs and GFP-HDFs. The original blots are presented in Supplementary Figs. 8, 9, 10, 11 and 12.

Article Snippet: After 72 h, each AAV2 vector was purified using the AAVpro® Purification Kit (AAV2) (Takara Bio).

Techniques: Expressing, Mutagenesis, Control, Western Blot, Phospho-proteomics, Infection, Activation Assay