control gfp  (Sino Biological)

Journal: bioRxiv

Article Title: Host DNA repair factors empower a mechanism of antiviral nucleoside analog resistance

doi: 10.64898/2026.05.21.726832

Figure Lengend Snippet: (A) Whole cell lysates from XPA - fibroblasts transduced with GFP-expressing control vector lentivirus (+GFP, left lane) or one expressing XPA-GFP (+XPA-GFP, right lane) were collected to determine XPA expression by immunoblotting. Tubulin serves as a loading control. (B) XPA - fibroblasts were infected at an MOI of 1 with WT or F412V. At 2 hpi, virus was washed out and media was replaced containing various concentrations of GCV. At four days post infection, cells and supernatant were collected for analysis by TCID 50 . Virus yield at each GCV concentration was compared to the DMSO control for each condition and plotted as percent yield reduction for three biological replicate experiments. Curves were similarly fitted for WT and F412V in each individual experiment to obtain ED 50 values and CIs for each condition. (C) Those ED 50 values from three biological replicate experiments were used for a ratio paired t test comparing viruses within each cell type and WT in the two cell types and the Holm-Šídák method was used to correct for multiple comparisons. ED 50 mean values are plotted with standard deviation error bars.

Article Snippet: For , to establish the XPA rescue and control cell lines, GM05509 fibroblasts ( XPA - ) were transduced in a 10 cm dish at MOI 3 with lentiviral particles to express XPA-GFP (Origene RC204872L2V) or unfused GFP control (Origene PS100093V) and 10 μg/mL polybrene.

Techniques: Transduction, Expressing, Control, Plasmid Preparation, Western Blot, Infection, Virus, Concentration Assay, Standard Deviation

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Experimental timeline. B Left: Maximum projection intensity images of an axon from cells co-expressing NC-GFP (scrambled negative control) and miRFP703-EB3 (far-red tagged EB3 protein). Red arrows ( B – E ) point to the base of the selected projection. Right: Selected axon and kymograph of miRFP703-EB3. For all kymographs ( B – E ), the vertical arrow represents distance, with the base of the arrow positioned towards the soma and the arrowhead positioned towards the tip of the projection. The horizontal arrows represent time progressing from left to right. C Left: Maximum projection intensity images of axons from cells co-expressing shKif11 (shRNA targeting KIF11) and miRFP703-EB3. Right: Selected axon and kymograph of miRFP703-EB3. D Left: Maximum projection intensity images of dendrites from cells co-expressing NC-GFP (scrambled negative control) and miRFP703-EB3. Right: Selected dendrite and kymograph of miRFP703-EB3. E Left: Maximum projection intensity images of dendrites from cells co-expressing shKif11 (shRNA targeting KIF11) and miRFP703-EB3. Right: Selected dendrite and kymograph of miRFP703-EB3.The percentage of minus-end-out MTs in axons ( F ) and dendrites ( G ) in NC-GFP or shKIF11 neurons. Two-tailed Unpaired t-test. EB3-comet flux in axons ( H ) and dendrites ( I ) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s test. EB3-comet growth rate in axons ( J ) and dendrites ( K ) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s test. EB3-comet distance traveled (MT growth) in axons ( L ) and dendrites ( M ) in NC-GFP or shKIF11 neurons. One-way ANOVA, Tukey’s multiple comparison test. N The percentage of minus-end-out MTs in primary, secondary, and tertiary dendrites. Mixed-effects model (REML) followed by Tukey’s test. O EB3-comet flux for plus-end-out and minus-end-out EB3 comets in secondary dendrites. One-way ANOVA, Tukey’s test. For all graphs ( F–O ), error bars represent ±SEM. P -values are listed above respective comparisons. For F , H , J , L , N = 10, 16 neurons and axons for NC-GFP or shKIF11, respectively. For G , I , K , M , N = 16(40), 15(29) neurons (dendrites) for NC-GFP and shKIF11, respectively. For O and N , NC-GFP N = 16 neurons (4 primary, 14 secondary, and 14 tertiary dendrites), shKIF11 N = 15 neurons (4 primary, 16 secondary, and 9 tertiary dendrites). Source data are provided as a file.

Article Snippet: DIV14-16 Primary hippocampal mouse neurons, plated in 35 mm Mattek No1.5 dishes were simultaneously transfected via combiMag and Lipofectamine with 0.5 μg EB3-miRFP703 (Addgene #79994) or mRuby-Synaptophysin [(pEF Synaptophysin-mRuby was a gift from Edwin Chapman (Addgene plasmid # 188980; http://n2t.net/addgene:188980 ; RRID:Addgene_188980)] or mApple-PSD95 [mApple-PSD95-N-14 was a gift from Michael Davidson (Addgene plasmid # 54941; http://n2t.net/addgene:54941 ; RRID:Addgene_54941)] and either 0.5 μg NC-GFP (Origene TR30013) or KIF11-shRNA-A (Origene TG501174) or tagged KIF11 constructs.

Techniques: Expressing, Negative Control, shRNA, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Experimental timeline. B Left: Maximum projection intensity image from a neuron co-expressing NC-GFP (scrambled negative control) and mRuby-Synaptophysin. Right: Selected axon and kymograph of mRuby-Synaptophysin. For all kymographs ( B , C , I , J ), the vertical arrow represents distance, with the base of the arrow positioned towards the soma and the arrowhead positioned towards the tip of the projection. The horizontal arrows represent time progressing from left to right. C Left: Maximum projection intensity image from a neuron co-expressing shKif11 (shRNA targeting KIF11) and mRuby-Synaptophysin. Right: Selected axon and kymograph of mRuby-Synaptophysin. D . Percentage of mobile mRuby-Synaptophysin in NC-GFP and shKIF11 primary hippocampal neurons’ axons. Two-tailed Unpaired t-test. In D–G N = 11, 12 neurons/axons for NC-GFP and shKif11, respectively. The flux of mRuby-Synaptophysin ( E ), the velocity of mRuby-Synaptophysin ( F ), and the distance traveled of mRuby-Synaptophysin ( G ) in NC-GFP and shKIF11 primary hippocampal neurons. One-way ANOVA, Tukey’s multiple comparison test. H . Experimental timeline. I Left: Maximum projection intensity images of dendrites from cells co-expressing NC-GFP and mApple-PSD95. Right: Selected dendrite and kymograph of mApple-PSD95. J Left: Maximum projection intensity images of dendrites from cells co-expressing shKif11 and mApple-PSD95. Right: Selected dendrite and kymograph of mApple-PSD95. K Percentage of mobile mApple-PSD95 in NC-GFP and shKIF11 primary hippocampal neurons. Two-tailed Unpaired t test. mApple-PSD95 flux ( L ), mApple-PSD95 velocity ( M ), and mApple-PSD95 distance traveled ( N ) in NC-GFP and shKIF11 primary hippocampal neurons. One-way ANOVA. Tukey’s multiple comparison test. In K–N N = 13, 9 neurons/dendrites for NC-GFP and shKif11, respectively. For all figures, white arrowheads point to the base of the selected axon/dendrites, red arrowheads highlight trafficking synaptophysin/PDS95. For all graphs ( D–G , K–N ), error bars represent ±SEM. P -values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV14-16 Primary hippocampal mouse neurons, plated in 35 mm Mattek No1.5 dishes were simultaneously transfected via combiMag and Lipofectamine with 0.5 μg EB3-miRFP703 (Addgene #79994) or mRuby-Synaptophysin [(pEF Synaptophysin-mRuby was a gift from Edwin Chapman (Addgene plasmid # 188980; http://n2t.net/addgene:188980 ; RRID:Addgene_188980)] or mApple-PSD95 [mApple-PSD95-N-14 was a gift from Michael Davidson (Addgene plasmid # 54941; http://n2t.net/addgene:54941 ; RRID:Addgene_54941)] and either 0.5 μg NC-GFP (Origene TR30013) or KIF11-shRNA-A (Origene TG501174) or tagged KIF11 constructs.

Techniques: Expressing, Negative Control, shRNA, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Schema of selected Microcephaly with or without chorioretinopathy, lymphedema, or intellectual disabilities (MCLID) patient mutations ( Hs: Homo sapiens ) and the corresponding mouse homolog (Mm: Mus Musculus ) (adapted from Schlögel et al. ) on the KIF11 protein. B Experimental timeline. C Confocal projection images of primary hippocampal mouse neurons transfected with control or KIF11 constructs, with the soma in the center of the image. Scale Bar=25 µm. D Soma size quantification of ( C ). N = 20,22,22,19 neurons for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Tukey’s test. E Quantification of dendritic morphology changes using Sholl analysis. N = 17,20,20,16 neurons for NC-GFP, KIF11-OE, KIF11 Y81F, and KIF11 ΔCterm , respectively. Two-way ANOVA followed by Tukey’s test. F Plus-end-out EB3-comet flux in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. G . Minus-end-out EB3-comets flux in KIF11 dendrites in comparison to NC-GFP. N = 14,16,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. H Percentage of Minus-end-out EB3-comets in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,22 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. I . Length of plus-end-out MT growth for KIF11 dendrites in comparison to NC-GFP. N = 17(248),17(59),18(114),22(184) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. J Length of minus-end-out MT growth for KIF11 dendrites compared to NC-GFP. N = 17(70),17(42),18(27),22(45) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. K Plus-end-out MT growth-rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(254),17(56),18(130),22(189) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. L Minus-end-out MT growth rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(59),17(49),18(32),22(41) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. For all graphs ( D–L ), error bars represent ±SEM. P values are listed above respective comparisons. P values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV14-16 Primary hippocampal mouse neurons, plated in 35 mm Mattek No1.5 dishes were simultaneously transfected via combiMag and Lipofectamine with 0.5 μg EB3-miRFP703 (Addgene #79994) or mRuby-Synaptophysin [(pEF Synaptophysin-mRuby was a gift from Edwin Chapman (Addgene plasmid # 188980; http://n2t.net/addgene:188980 ; RRID:Addgene_188980)] or mApple-PSD95 [mApple-PSD95-N-14 was a gift from Michael Davidson (Addgene plasmid # 54941; http://n2t.net/addgene:54941 ; RRID:Addgene_54941)] and either 0.5 μg NC-GFP (Origene TR30013) or KIF11-shRNA-A (Origene TG501174) or tagged KIF11 constructs.

Techniques: Transfection, Control, Construct, Comparison

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Timeline of experimental design to record miniature excitatory post-synaptic potential (mEPSCs) in mouse primary hippocampal culture expressing NC-GFP or KIF11 constructs. Tetrodotoxin (TTX) was added to ensure mEPSCs and not spontaneous EPSCs were captured. B Two representative traces of mEPSCs for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm 24–48 h post-transfection. Bar graph of mEPSC amplitude ( C ) and frequency ( D ) in NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons. N = 10,13,12,13 NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons respectively. One-way ANOVA followed by Tukey’s Multiple comparisons test. Cumulative probability graphs showing no change in mEPSC amplitude ( E ), but reduced frequency ( F ) in KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons compared to NC-GFP. Kolmogorov-Smirnov Test. For graphs ( C , D ), error bars represent ± SEM. P-values are listed above respective comparisons. Source data are provided as a Source Data file.

Article Snippet: DIV14-16 Primary hippocampal mouse neurons, plated in 35 mm Mattek No1.5 dishes were simultaneously transfected via combiMag and Lipofectamine with 0.5 μg EB3-miRFP703 (Addgene #79994) or mRuby-Synaptophysin [(pEF Synaptophysin-mRuby was a gift from Edwin Chapman (Addgene plasmid # 188980; http://n2t.net/addgene:188980 ; RRID:Addgene_188980)] or mApple-PSD95 [mApple-PSD95-N-14 was a gift from Michael Davidson (Addgene plasmid # 54941; http://n2t.net/addgene:54941 ; RRID:Addgene_54941)] and either 0.5 μg NC-GFP (Origene TR30013) or KIF11-shRNA-A (Origene TG501174) or tagged KIF11 constructs.

Techniques: Expressing, Construct, Transfection