lentiviral constructs  (Addgene inc)

Journal: Nature Aging

Article Title: Targeting iron-associated protein Ftl1 in the brain of old mice improves age-related cognitive impairment

doi: 10.1038/s43587-025-00940-z

Figure Lengend Snippet: Primary neurons were infected with lentivirus encoding Ftl1 or control at 8 days in vitro (DIV8) and analyzed at DIV18. ( a ) Illustration showing the coverslip regions used for neurite analysis of primary neurons following lentiviral-mediated overexpression of Ftl1 (Ftl1 OE) or control. ( b ) Representative images of GFP-positive neurons used as a redout for lentiviral infection for both Ftl1 OE and control conditions. ( c ) Representative Western blot and quantification of FTL1 protein levels N = 8 replicates/condition (p-value < 0.0001). ( d ) Quantification of cell death in primary neurons following viral-mediated overexpression of Ftl1 by LDH assay. N = 5 positive control, N = 5 control and N = 5 Ftl1 OE replicates/condition (p-value < 0.0001). ( e ) Representative images and quantification of DAPI-positive nuclei of Ftl1 OE and control neurons at DIV18. N = 5 controls and N = 5 Ftl1 OE replicates/condition (p-value = 0.8956). Data shown as mean±s.e.m.; ****P < 0.001, two-tailed t-test ( c,d,e ).

Article Snippet: Validated gRNA sequences were subcloned into a Syn1 -Cre lentiviral vector (Addgene, no. 68841; ref. ) using PacI and NheI/XbaI.

Techniques: Infection, Control, In Vitro, Over Expression, Western Blot, Lactate Dehydrogenase Assay, Positive Control, Two Tailed Test

Journal: Nature Aging

Article Title: Targeting iron-associated protein Ftl1 in the brain of old mice improves age-related cognitive impairment

doi: 10.1038/s43587-025-00940-z

Figure Lengend Snippet: Primary neurons were infected with lentivirus encoding shRNA targeting Ftl1 or control at 8 days in vitro (DIV8) and analyzed at DIV18. ( a ) Illustration showing the coverslip regions used for neurite analysis of primary neurons following lentiviral-mediated Ftl1 knockdown (Ftl1 KD) or control. ( b ) Representative images of GFP-positive neurons used as a redout for lentiviral infection for both Ftl1 KD and control conditions. ( c ) Representative Western blot and quantification of FTL1 protein levels. N = 8 replicates/condition N= (p-value < 0.0001) ( d ) Quantification of cell death in primary neurons following viral-mediated overexpression of Ftl1 by LDH assay. ( d ) Representative images and quantification of DAPI-positive nuclei of Ftl1 KD and control neurons at DIV18. N = 5 positive control, N = 5 control and N = 5 Ftl1 KD replicates/condition (p-value < 0.0001). ( e ) Representative images and quantification of DAPI-positive nuclei of Ftl1 KD and control neurons at DIV18. N = 5 controls and N = 5 Ftl1 KD replicates/condition (p-value = 0.2130). Data shown as mean±s.e.m.; ****P < 0.001, two-tailed t-test ( c,d,e ).

Article Snippet: Validated gRNA sequences were subcloned into a Syn1 -Cre lentiviral vector (Addgene, no. 68841; ref. ) using PacI and NheI/XbaI.

Techniques: Infection, shRNA, Control, In Vitro, Knockdown, Western Blot, Over Expression, Lactate Dehydrogenase Assay, Positive Control, Two Tailed Test

Journal: Nature Aging

Article Title: Targeting iron-associated protein Ftl1 in the brain of old mice improves age-related cognitive impairment

doi: 10.1038/s43587-025-00940-z

Figure Lengend Snippet: Primary neurons were infected with lentivirus encoding Ftl1 or control at 8 days in vitro (DIV8) subsequently treated with NADH (200uM) for 5 consecutive days starting at DIV14 and analyzed at DIV18. ( a ) Schematic illustrating experimental timeline for lentiviral-mediated overexpression of Ftl1 (Ftl1 OE) or control and NADH treatment in primary neurons. ( b ) Quantification of cell death in primary neurons following viral-mediated overexpression of Ftl1 and NADH treatment by Lactate Dyhydrogenase (LDH) assay. Each data point represents the average of 3 replicates. Data shown as mean±s.e.m, two-way ANOVA followed by Tukey’s post hoc t-test.

Article Snippet: Validated gRNA sequences were subcloned into a Syn1 -Cre lentiviral vector (Addgene, no. 68841; ref. ) using PacI and NheI/XbaI.

Techniques: Infection, Control, In Vitro, Over Expression, Lactate Dehydrogenase Assay