Journal: bioRxiv

Article Title: Pyruvate promotes ciliogenesis bypassing IFT88 dependency and attenuates DSS-induced colitis

doi: 10.64898/2025.12.17.694572

Figure Lengend Snippet: PC length (A, C, E) and number of PC-positive cells (B, D, F) in 24h serum starved CF cultures in absence or presence of 2mM sodium pyruvate and supplemented with indicated concentrations of MEK1/2 inhibitor PD9805 (A, B), ERK1/2 inhibitor SCH772984 ( C, D) and histone acetylation EP300 inhibitor C646 ( E, F). Statistical analysis: *p<0.05, **p< 0.01, ***p< 0.001, ****p<0.0001 by two-tailed unpaired t-test.

Article Snippet: The MEK1/2 inhibitor PD98059 (used at 20μM), ERK1/2 inhibitor SCH772984 (used at 300nM), the inhibitor of pyruvate transport at the plasma membrane AZD3965 (used at 200nM), the microtubule acetylation inhibitor GM-90257 (used at 500nM) and the histone acetyl transferase inhibitors for E1A-associated Protein p300 (Ep300) C646 (used at 25μM) were purchased from MCE (MedChem Express, Monmouth Junction, NJ, USA).

Techniques: Two Tailed Test

Journal: bioRxiv

Article Title: A single-component optogenetic toolkit for programmable control of microtubule

doi: 10.1101/2025.10.31.685931

Figure Lengend Snippet: (a) Domain organization of mouse KIF5A (mKIF5A), showing the N-terminal motor head, central coiled-coil stalk, and C-terminal cargo-binding tail. (b) Schematic of mKIF5A truncation variants fused to CRY2 for optogenetic activation. Variant V4 (residues 1-379) was identified as the optimal construct and designated OptoMotor. (c) Design principle of OptoMotor. Blue light illumination induces CRY2 oligomerization to drive reassembly of the truncated motor complex and restoring plus-end-directed motility along under MTs. (d) Confocal images of selected CRY2-KIF5A truncation variants expressed in HeLa cells under dark and illuminated conditions. Variant V1 exhibited constitutive peripheral accumulation; V4 showed robust light-induced redistribution to the cell periphery without appreciable basal activity; and V6 displayed pronounced MT labeling. See Supplementary Fig. 7 for the complete set of variants. (e) Quantification of periphery-to-cytosol fluorescence ratios across truncation variants. Each symbol represents the mean of 5-6 cells from a single imaging field. A total of 40-50 cells were analyzed per construct across three independent biological replicates. (f) Time-lapse images showing reversible distribution of OptoMotor during repeated dark-light cycles. Also see Supplementary Video 11 . (g) Kinetic trace of peripheral intensity changes showing OptoMotor activation (t 1/2 , ON = 0.9 min) and deactivation (t 1/2 , OFF = 3.7 min). (h) Localized blue light illumination within defined regions of interest (ROIs R1, R2) elicited spatially confined redistribution of OptoMotor in single cells. (i) Quantification of relative fluorescence intensity changes within ROIs confirming high spatial precision of light-triggered activation. (j) Schematic of the chimeric OptoMotor-Tail construct, generated by fusing the cargo-binding C-terminal tail of mKIF5A (residues 907-1027) to OptoMotor. Upon blue light activation, OptoMotor-Tail drives peripheral transport of lysosomes to enhance mTORC signaling. (k) Confocal images of HeLa cells co-expressing LAMP1-GFP and mCh-OptoMotor-Tail showing light-induced redistribution of lysosomes toward the cell periphery. (l) Immunoblot showing phosphorylation of S6K at T389 (P-S6K) as a readout of mTORC1 activity. Blue light illumination enhanced P-S6K levels in cells expressing OptoMotor-Tail, consistent with lysosomal repositioning mediated activation of mTORC1 signaling. Starved cells before and after nutrient recovery were used as negative and positive controls for reporting mTORC1 activity.

Article Snippet: The plasmid templates for EB1 (#17234), CLIP170 (#54044), CAMSAP1 (# 59036),CAMSAP2 (#59037), KIF5A (#166954), and spastin (#134461) were purchased from Addgene.

Techniques: Binding Assay, Activation Assay, Variant Assay, Construct, Activity Assay, Labeling, Fluorescence, Imaging, Generated, Expressing, Western Blot, Phospho-proteomics

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Flavonoid compound from Forsythia suspensa leaves inhibits adenovirus infection related to cell cycle based on UHPLC-Q-Exactive-Orbitrap/MS and experimental validation

doi: 10.3389/fcimb.2025.1627863

Figure Lengend Snippet: In vitro experiments validated the FSL inhibition of HAdV infection. A549 cell cultures were treated with varying concentrations of FSL prior to HAdV infection (MOI = 1) for 24 h. Real-time PCR and immunoblotting were used to detect HAdV protein or gene expression. (A–D) Real-time PCR was utilized to analyze the levels of the HAdV E1A, E2, E3, and E4 mRNA expression and normalized to GAPDH. (E, F) Immunoblotting was used to detect the expression of HAdV E1A proteins and GAPDH. The experimental procedures were independently replicated three times. The data are presented as mean ± SD ( n = 3) of one experiment. Statistical significance was denoted as ns, no significant, **** p <0.00001, ** p < 0.01, and *** p < 0.0001.

Article Snippet: The membranes were blocked by using 5% skimmed milk and incubated with Adenovirus-2/5 E1A antibody (1:1,000, Santa Cruz, sc-25, lot #K1022), CDC25A (1:1,000, Beyotime, AF6459), CCNB2 (1:1,000, zenbio, R381384), AURKA (1:1,000, Zenbio, 222073), CHEK1 (1:1,000, Beyotime, AF6498), and CCNA2 (1:1,000, Beyotime, AF6624), with GAPDH (1:2,000, Beyotime, AF0006) serving as the loading control.

Techniques: In Vitro, Inhibition, Infection, Real-time Polymerase Chain Reaction, Western Blot, Gene Expression, Expressing