Journal: Molecular Biology of the Cell

Article Title: Microtubule motors involved in nuclear movement during skeletal muscle differentiation

doi: 10.1091/mbc.E16-06-0405

Figure Lengend Snippet: siRNA sequences and Taqman probes used for each molecular motor.

Article Snippet: NM_001025360 , Klc1 , GGAGGAGAAGAAACACCUGtt , CAGGUGUUUCUUCUCCUCCtc , Mm00492936_m1.

Techniques: Sequencing, TaqMan Assay

Journal: Translational Lung Cancer Research

Article Title: Endoplasmic reticulum stress-related genes contribute to lung cancer risk: a multiomics data integration study

doi: 10.21037/tlcr-2025-474

Figure Lengend Snippet: Integrated multiomics analysis of KLC1 . (A) SMR analysis results from the integration of lung cancer GWAS and mQTL data at the KLC1 locus. (B) SMR analysis results of lung cancer GWAS and eQTL at the KLC1 locus. (C) SMR analysis results of lung cancer GWAS and pQTL at the KLC1 locus. (D) Correlation of SNP effect sizes for KLC1 between lung cancer GWAS and QTL data (mQTL, eQTL, pQTL). cis-QTL, cis-acting quantitative trait loci; eQTL, expression quantitative trait loci; GWAS, genome-wide association study; mQTL, methylation quantitative trait loci; pQTL, protein quantitative trait loci; SMR, summary data-based Mendelian randomization; SNP, single-nucleotide polymorphism.

Article Snippet: The antibodies employed in this IHC analysis were BCL2L1 (bs-1336R; Bioss Antibodies, Woburn, MA, USA) and KLC1 (bs-11212R; Bioss Antibodies).

Techniques: Expressing, GWAS, Methylation

Journal: Translational Lung Cancer Research

Article Title: Endoplasmic reticulum stress-related genes contribute to lung cancer risk: a multiomics data integration study

doi: 10.21037/tlcr-2025-474

Figure Lengend Snippet: Integrated multiomics analysis of BCL2L1 . (A) SMR analysis results from the integration of lung cancer GWAS and mQTL data at the BCL2L1 locus. (B) SMR analysis results of lung cancer GWAS and eQTL data at the BCL2L1 locus. (C) Correlation of SNP effect sizes for BCL2L1 between lung cancer GWAS and QTL data (mQTL, eQTL). (D) Manhattan plots displaying the SMR results for KLC1 and BCL2L1 at the multiomics level. CHR on the X-axis indicates chromosome numbers. cis-QTL, cis-acting quantitative trait loci; eQTL, expression quantitative trait loci; GWAS, genome-wide association study; mQTL, methylation quantitative trait loci; pQTL, protein quantitative trait loci; SNP, single-nucleotide polymorphism; SMR, summary data-based Mendelian randomization.

Article Snippet: The antibodies employed in this IHC analysis were BCL2L1 (bs-1336R; Bioss Antibodies, Woburn, MA, USA) and KLC1 (bs-11212R; Bioss Antibodies).

Techniques: Expressing, GWAS, Methylation

Journal: Translational Lung Cancer Research

Article Title: Endoplasmic reticulum stress-related genes contribute to lung cancer risk: a multiomics data integration study

doi: 10.21037/tlcr-2025-474

Figure Lengend Snippet: Expression and prognostic analysis of BCL2L1 and KCL1 . (A) Differential expression analysis of KLC1 and BCL2L1 between lung cancer and normal samples. (B) Comparison of immune cell infiltration levels in the lung cancer microenvironment between high- and low-expression groups for KLC1 and BCL2L1. (C) Overall survival analysis comparing high- and low-expression groups of KLC1 and BCL2L1. (D) Immunohistochemical analyses of KLC1 in lung cancer tissue from the HPA database. ( https://www.proteinatlas.org/ENSG00000126214-KLC1/cancer/lung+cancer ). (E) Immunohistochemical staining of BCL2L1 and KLC1 in lung cancer and normal lung tissues (scale bar: 50 µm). *, P<0.05; **, P<0.01; ***, P<0.001. CI, confidence interval; HPA, Human Protein Atlas; HR, hazard ratio; CI, confident interval; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; N, normal lung tissues; T, lung cancer tissues; TPM, transcripts per million.

Article Snippet: The antibodies employed in this IHC analysis were BCL2L1 (bs-1336R; Bioss Antibodies, Woburn, MA, USA) and KLC1 (bs-11212R; Bioss Antibodies).

Techniques: Expressing, Quantitative Proteomics, Comparison, Immunohistochemical staining, Staining

Journal: Translational Lung Cancer Research

Article Title: Endoplasmic reticulum stress-related genes contribute to lung cancer risk: a multiomics data integration study

doi: 10.21037/tlcr-2025-474

Figure Lengend Snippet: Diagram illustrating the regulatory mechanisms and functions of the ERS-related genes KLC1 and BCL2L1 in lung cancer. ERS, endoplasmic reticulum stress.

Article Snippet: The antibodies employed in this IHC analysis were BCL2L1 (bs-1336R; Bioss Antibodies, Woburn, MA, USA) and KLC1 (bs-11212R; Bioss Antibodies).

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Kinesin light chain-1 serine-460 phosphorylation is altered in Alzheimer’s disease and regulates axonal transport and processing of the amyloid precursor protein

doi: 10.1186/s40478-019-0857-5

Figure Lengend Snippet: Characterisation of KLC1 serine-460 phospho-specific antibody. HEK293 cells were transfected with either control vector, FLAG-KLC1S460A or FLAG-KLC1wt and treated with either vehicle or okadaic acid (OA) as indicated. The different samples were then probed on immunoblots with antibodies to serine-460 phosphorylated KLC1 (KLC1-ser460p), FLAG to detect total KLC1 (FLAG-KLC1), active and total ERK1/2, and tubulin. The KLC1 serine-460 phospho-specific antibody detects KLC1wt but not KLC1S460A and signals with the antibody are increased in KLC1wt cells treated with okadaic acid that activates the KLC1 serine-460 kinase ERK1/2

Article Snippet: Rabbit antibody to KLC1 phosphorylated on serine-460 was generated by immunisation with keyhole limpet hemocyanin coupled to peptide CKVDS phos PTVTTTLKNL in which serine-460 (S phos ) was phosphorylated (Proteintech).

Techniques: Transfection, Control, Plasmid Preparation, Western Blot

Journal: Acta Neuropathologica Communications

Article Title: Kinesin light chain-1 serine-460 phosphorylation is altered in Alzheimer’s disease and regulates axonal transport and processing of the amyloid precursor protein

doi: 10.1186/s40478-019-0857-5

Figure Lengend Snippet: KLC1 levels are reduced and the relative levels of KLC1 serine-460 phosphorylation are increased in Alzheimer’s disease frontal cortex. Representative immunoblots showing total KLC1, KLC1 serine-460 phosphorylation (KLC1S460p) and NSE levels in post-mortem human control (Ctrl) and Alzheimer’s disease frontal cortex. Braak stages are indicated. Graphs show quantification of total KLC1 and KLC1 phosphorylated on serine-460 in the different samples. KLC1 and KLC1S460p signals were normalised to NSE levels. These normalised KLC1 levels were then used to quantify changes to total KLC1 and KLC1S460p (expressed as the ratio of KLC1S460p/total KLC1). Data were analysed by Welch’s ANOVA and Games-Howell post hoc test. N = 13–15, error bars are s.e.m., * p < 0.05 ** p < 0.01, *** p < 0.001, ns not significant

Article Snippet: Rabbit antibody to KLC1 phosphorylated on serine-460 was generated by immunisation with keyhole limpet hemocyanin coupled to peptide CKVDS phos PTVTTTLKNL in which serine-460 (S phos ) was phosphorylated (Proteintech).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Acta Neuropathologica Communications

Article Title: Kinesin light chain-1 serine-460 phosphorylation is altered in Alzheimer’s disease and regulates axonal transport and processing of the amyloid precursor protein

doi: 10.1186/s40478-019-0857-5

Figure Lengend Snippet: Mutation of KLC1 serine-460 to mimic permanent phosphorylation, inhibits axonal transport of APP in cultured rat cortical neurons. ( a ) Representative kymographs showing axonal transport of APP-EGFP in APP-EGFP+KLC1wt and APP-EGFP+KLC1S460D co-transfected neurons; scale bar and times are indicated. Bar charts show % for total, anterograde and retrograde APP-EGFP movement. N = 17 EGFP+KLC1wt and 20 APP-EGFP+KLC1S460D co-transfected neurons. Statistical significance was determined by Student’s t-test. Error bars are s.e.m.; ** p < 0.01; ns not significant. ( b ) Violin plots show velocities of APP-EGFP movement in anterograde and retrograde directions in the different transfected cells. Median and interquartile ranges are indicated by hashed lines. N = 163 anterogradely and 62 retrogradely moving APP-EGFP in KLC1wt and 179 anterogradely and 54 retrogradely moving APP-EGFP in KLC1S460D co-transfected cells. Statistical significance was determined by Mann-Whitney U test; ns not significant

Article Snippet: Rabbit antibody to KLC1 phosphorylated on serine-460 was generated by immunisation with keyhole limpet hemocyanin coupled to peptide CKVDS phos PTVTTTLKNL in which serine-460 (S phos ) was phosphorylated (Proteintech).

Techniques: Mutagenesis, Phospho-proteomics, Cell Culture, Transfection, MANN-WHITNEY

Journal: Acta Neuropathologica Communications

Article Title: Kinesin light chain-1 serine-460 phosphorylation is altered in Alzheimer’s disease and regulates axonal transport and processing of the amyloid precursor protein

doi: 10.1186/s40478-019-0857-5

Figure Lengend Snippet: Mutation of endogenous Drosophila KLC serine-433 to mimic permanent phosphorylation inhibits axonal transport of APP in wing sensory neurons. ( a ) Alignment showing high conservation of the amino acid sequences encompassing rat KLC1 serine-460 and the homologous Drosophila KLC serine-433 (indicated in red). ( b ) CRISPR genome editing approach to mutate KLC serine-433 to aspartic acid. Drosophila Klc sequence (upper line) along with the ssODN (lower line) are shown. Serine-433 codon (KLC) and aspartic acid codon (ssODN) are shown in red, the protospacer sequence in grey shade and the PAM site in blue shade. A G-to-A transition is introduced to mutate the PAM. ( c ) Representative kymographs showing axonal transport of APP-YFP in Klcwt and KlcS433D homozygous backgrounds in 2-day old Drosophila ; scale bar and times are indicated. Bar chart shows the relative proportions of stationary, anterograde and retrograde moving APP-YFP in the Klcwt background. ( d ) Bar charts show total, anterograde and retrograde APP-YFP movement runs in Klcwt and KlcS433D backgrounds. ( e ) Violin plots show velocities of APP-YFP runs in anterograde and retrograde directions in the different backgrounds. Median and interquartile ranges are indicated by hashed lines. N = 8 wings for each genotype. For velocity studies, N = 577 anterogradely and 347 retrogradely moving APP-YFP in Klcwt , and 343 anterogradely and 247 retrogradely moving APP-YFP in the KlcS433D background. Statistical significance was determined by one-way ANOVA with Holm-Sidak’s multiple comparison test in ( c ), Mann-Whitney U test in ( d ) and two-tailed Student’s t test in ( e ). Error bars are s.e.m.; **p < 0.01; **** p < 0.0001; ns not significant

Article Snippet: Rabbit antibody to KLC1 phosphorylated on serine-460 was generated by immunisation with keyhole limpet hemocyanin coupled to peptide CKVDS phos PTVTTTLKNL in which serine-460 (S phos ) was phosphorylated (Proteintech).

Techniques: Mutagenesis, Phospho-proteomics, CRISPR, Sequencing, Comparison, MANN-WHITNEY, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Kinesin light chain-1 serine-460 phosphorylation is altered in Alzheimer’s disease and regulates axonal transport and processing of the amyloid precursor protein

doi: 10.1186/s40478-019-0857-5

Figure Lengend Snippet: Expression of KLC1 serine-460 to mimic permanent phosphorylation promotes amyloidogenic processing of APP. ( a ) APP processing was monitored in HEK293 cells essentially as described using an APP-GAL4 reporter assay that drives GAL4-UAS-dependent expression of firefly luciferase . Luciferase signals were normalised to co-transfected Renilla signals to correct for transfection efficiency. Bar chart shows relative luciferase signals in cells transfected with the GAL4-UAS-luciferase+Renilla reporters plus either APP-GAL4 + KLC1wt or APP-GAL4 + KLC1S460D as indicated. N = 15 from 3 independent experiments. Data were analysed by Student’s t test, error bars are s.e.m., * p < 0.05. ( b ) Expression of KLC1S460D increases Aβ production. Levels of Aβ (1–38), Aβ (1–40) and Aβ (1–42) we quantified in conditioned media from cells co-transfected with APP+KLC1wt or APP+ KLC1S460D. N = 7; data were analysed by Student’s t test, error bars are s.e.m., * p < 0.05

Article Snippet: Rabbit antibody to KLC1 phosphorylated on serine-460 was generated by immunisation with keyhole limpet hemocyanin coupled to peptide CKVDS phos PTVTTTLKNL in which serine-460 (S phos ) was phosphorylated (Proteintech).

Techniques: Expressing, Phospho-proteomics, Reporter Assay, Luciferase, Transfection

Journal: The American Journal of Pathology

Article Title: A Unique Role for Endothelial Cell Kinesin Light Chain 1, Variant 1 in Leukocyte Transendothelial Migration

doi: 10.1016/j.ajpath.2016.01.011

Figure Lengend Snippet: Knockdown of kinesin-1 in HUVECs results in a decrease in LBRC targeted recycling. HUVECs were transduced with scrambled shRNA, kinesin-1 shRNA (KHC shRNA), or kinesin-1 shRNA with addition of the kinesin-1 rescue construct (KHC shRNA + Rescue). Monocytes were allowed to transmigrate for 8.5 minutes. A: Confocal stacks were imaged with CD18 in green and recycled LBRC in red. Orthogonal projections (xz) are depicted as smaller images to the right of their corresponding images and indicate whether the monocyte is in the process of TEM or engaging the endothelial cell border. Arrowheads indicate site of leukocyte TEM. Dotted lines in the orthogonal projection indicate abluminal surface of endothelial cells. Constitutive recycling occurs, including under the blocked leukocyte,5, 6 when kinesin-1 is knocked down, but there is no enrichment of LBRC around the monocyte. The KHC shRNA + Rescue panel shows two transmigrating monocytes, one exhibiting a ring of enrichment surrounding it and the other exhibiting local enrichment at both sides of it. B: LBRC enrichment was measured around all adherent monocytes. The dotted line represents no change in LBRC enrichment relative to adjacent junctional LBRC intensity. C: Targeted recycling significantly decreased on knockdown of KLC1 and was restored to control levels with the rescue. Data are expressed as means ± SEM. n = 3 experiments with three monolayers per condition per experiment and >300 monocyte/endothelial cell interactions per experiment (B and C). ∗∗∗P < 0.001. Scale bar = 10 μm. HUVEC, human umbilical vein endothelial cell; KHC, kinesin heavy chain; LBRC, lateral border recycling compartment; PBMC, peripheral blood mononuclear cell; TEM, transendothelial migration.

Article Snippet: The KLC1C was rescued by KLC1 variant 1 cDNA (ViGene Biosciences, Inc., Rockville, MD).

Techniques: Transduction, shRNA, Construct, Migration

Journal: The American Journal of Pathology

Article Title: A Unique Role for Endothelial Cell Kinesin Light Chain 1, Variant 1 in Leukocyte Transendothelial Migration

doi: 10.1016/j.ajpath.2016.01.011

Figure Lengend Snippet: Knockdown of KLC1 isoform variant 1 in HUVEC blocks TEM. A: Western blot analysis of KLC2 expression in HEK293 cells versus HUVECs demonstrates absence of KLC2 in HUVECs. Cells were plated on Mattek dishes and lyzed 96 hours later. The amount of lysate added to the gels is indicated and was adjusted to obtain similar KLC1 expression levels in 293 cells versus HUVECs to fairly assess KLC2 levels in 293 cells versus HUVEC. B: HUVECs were transduced with scrambled shRNA, KLC1 variant 1 shRNA, KLC1 variants 2 and 3 shRNA, or KLC1 variant 1 shRNA and the KLC1 variant 1 rescue construct. Cells were lyzed 72 hours later. Quantification of the Western blot analysis. C and D: HUVECs were transduced with scrambled shRNA, KLC1 variant 1 shRNA, KLC1 variants 2 and 3 shRNA, KLC variants 1 to 3 shRNA, or KLC1 variant 1 shRNA with the KLC1 variant 1 rescue construct. Adhesion (C) and migration (D) to cell borders were measured after an 8.5-minute TEM period. Monocytes (E) or neutrophils (F) were allowed to transmigrate for 60 minutes before TEM was quantified. Data are expressed as means ± SEM. n = 3 experiments (B); n = 3 experiments with three monolayers per condition per experiment (C–F). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. HUVEC, human umbilical vein endothelial cell; KLC, kinesin light chain; PBMC, peripheral blood mononuclear cell; TEM, transendothelial migration.

Article Snippet: The KLC1C was rescued by KLC1 variant 1 cDNA (ViGene Biosciences, Inc., Rockville, MD).

Techniques: Variant Assay, Western Blot, Expressing, Transduction, shRNA, Construct, Migration

Journal: The American Journal of Pathology

Article Title: A Unique Role for Endothelial Cell Kinesin Light Chain 1, Variant 1 in Leukocyte Transendothelial Migration

doi: 10.1016/j.ajpath.2016.01.011

Figure Lengend Snippet: Knockdown of KLC1 isoform variant 1 in HUVECs results in a decrease in LBRC targeted recycling. HUVECs were transduced with scrambled shRNA, KLC1 variant 1 shRNA, KLC1 variants 2 and 3 shRNA, and KLC variants 1 to 3 shRNA. A and B: Monocytes were allowed to transmigrate for 8.5 minutes. Confocal stacks were imaged (A), with their corresponding orthogonal projections (xz) as smaller images to the right, and LBRC enrichment was measured around all adherent monocytes (B). The dotted line represents no change in LBRC enrichment. C: Targeted recycling significantly decreased on knockdown of KLC1 isoform variant 1. Arrowheads indicate site of leukocyte transendothelial migration. Dotted lines in the orthogonal projection indicate abluminal surface of endothelial cells. Data are expressed as means ± SEM. n = 3 experiments with three monolayers per condition per experiment (B and C). ∗∗∗P < 0.001. Scale bar = 10 μm. HUVEC, human umbilical vein endothelial cell; KLC, kinesin light chain; LBRC, lateral border recycling compartment; PBMC, peripheral blood mononuclear cell.

Article Snippet: The KLC1C was rescued by KLC1 variant 1 cDNA (ViGene Biosciences, Inc., Rockville, MD).

Techniques: Variant Assay, Transduction, shRNA, Migration