phalloidin buffer  (Santa Cruz Biotechnology)

Journal: The Journal of Cell Biology

Article Title: Energy stress activates AMPK to arrest mitochondria via phosphorylation of TRAK1

doi: 10.1083/jcb.202501023

Figure Lengend Snippet: Controls for rapalog motility experiments and perimitochondrial actin recruitment. (A) Representative example neurites showing rapalog recruits mCherry-FRB to mitochondria in the presence of vehicle or AntA. Neurons were transfected with mito-GFP-FKBP and mCherry-FRB and pretreated for 1 h with either 4 nM AntA or vehicle before addition of 1 µM rapalog as in . (B) Representative images of hippocampal neurons that were fixed and stained with Alexa Fluor 488 phalloidin conjugate and Hoechst. Either control neurons or neurons pretreated with 4 nM AntA were exposed to 5 µM LatA. Note the persistence of a LatA-resistant pool of actin after pretreatment with AntA. (C) Quantification of filamentous actin from the average Alexa Fluor 488 phalloidin intensity of 60× imaging fields of neurons treated as in B. (D) Quantification of mitochondrial motility in hippocampal neurons pretreated for 1 h with 4 nM AntA and then 30 min with both 5 uM LatA and 4 nM AntA. Once arrested by treatment with AntA, LatA could not reverse the arrest. (E) Time-lapse series of fibroblasts treated for 1 h with 40 nM AntA and imaged as in . The arrowhead indicates a mitochondrion that appears to lack perimitochondrial actin and moves, while nearby mitochondria do not. (F) Representative images of rat embryonic fibroblasts transduced with GFP-F-tractin and mito-mRaspberry from the quantification in . Cells cultured in galactose were treated with 40 nM AntA or vehicle. One hour after drug treatment, cells either had glucose or galactose added to a concentration of 25 mM, or no further addition. Cells were imaged after 30 min. P values for C were calculated by two-tailed, unpaired t tests with Welch’s correction. A blocked one-way ANOVA was performed with Tukey’s multiple comparison correction for D. Select comparisons are shown for D. For supplemental images, 3A scale bars = 10 µm. The scale bar in 3E is 5 µm. For supplemental images, 3B and 3F scale bars = 20 μm.

Article Snippet: The phalloidin buffer was washed off, and the sample was incubated for 30 min with AF488-conjugated phalloidin (sc-36379; Santa Cruz) dissolved in 1:1,000 in phalloidin buffer.

Techniques: Transfection, Staining, Control, Imaging, Transduction, Cell Culture, Concentration Assay, Two Tailed Test, Comparison

Journal: bioRxiv

Article Title: Unravelling the distinct phenotype and mechanosensitive properties of different tendon cell populations

doi: 10.64898/2026.01.08.698354

Figure Lengend Snippet: a) Schematic and photographic overview of the rat tail anatomy highlighting the location of interfascicular matrix (IFM) and fascicular matrix (FM) regions within the tendon, followed by a 3D schematic of bundle structure. b) Immunofluorescence of a rat tail quadrant (longitudinal cuts) stained with DAPI (nucleus) and phalloidin (actin cytoskeleton). IFM and FM regions are visually evident with different morphology and organisation of IFM and FM cells. c) The schematic depicts a transverse plane through an isolated fascicle surrounded by IFM as on removal from a tail tendon. Confocal images through the fascicle at different heights (shown on the left) demonstrate that it is feasible to detect when imaging within the IFM or FM region. Fascicles were digested in collagenase for 0 – 120 minutes, fixed, stained and imaged, and the thickness of the remaining IFM recorded at different time intervals, confirmed that IFM was consistently fully digested after 120 minutes. d) The digestion protocol was selected based on these data, discarding the digested cells collected between 90 minutes and 150 minutes to ensure IFM and FM digestions remained distinct for each cell population (n=3 rats, 2 fascicles/rat).

Article Snippet: Blocking was performed using a 1% BSA solution for 1 hour, before cells were stained with phalloidin (Texas RedTM-X Phalloidin ThermoFisher diluted in 1% BSA solution) for an additional 45 minutes, followed by three PBS rinses.

Techniques: Immunofluorescence, Staining, Isolation, Imaging

Journal: bioRxiv

Article Title: Unravelling the distinct phenotype and mechanosensitive properties of different tendon cell populations

doi: 10.64898/2026.01.08.698354

Figure Lengend Snippet: a) IFM and FM cells fixed and stained with DAPI (nucleus) and phalloidin (actin), comparing cell morphology at day 2 of P1 and P3. b) Comparison of mean cell area, cell nucleus area and actin expression/cell at P0, P2 and P3 (N=200/group, mean ± S.E.), comparing IFM cells (green) and FM cells (red). ****p<0.001. Further associated data is provided in .

Article Snippet: Blocking was performed using a 1% BSA solution for 1 hour, before cells were stained with phalloidin (Texas RedTM-X Phalloidin ThermoFisher diluted in 1% BSA solution) for an additional 45 minutes, followed by three PBS rinses.

Techniques: Staining, Comparison, Expressing

Journal: bioRxiv

Article Title: Unravelling the distinct phenotype and mechanosensitive properties of different tendon cell populations

doi: 10.64898/2026.01.08.698354

Figure Lengend Snippet: a) Typical images of IFM and FM cells at day 2 of P3 culture comparing collagen-coated TCP with 900 kPa and 20kPa surfaces, illustrating the morphological recovery. Cells are stained with DAPI (nucleus; blue) and Phalloidin (cytoskeleton, red). b) Image semi-quantification of changes in cell body area, nucleus area, and actin expression/cell at P3 of culture on substrates of varying stiffness. Mean ± S.E. of >200 cells/condition are displayed for IFM cells (green) and FM cells (red). *= p<0.001 highlights a significant difference between the condition and the P0 TCP reference. A more detailed breakdown of the distribution of cell morphologies for each condition and correlations between cell body area, nucleus area and actin expression/cell are shown in & .

Article Snippet: Blocking was performed using a 1% BSA solution for 1 hour, before cells were stained with phalloidin (Texas RedTM-X Phalloidin ThermoFisher diluted in 1% BSA solution) for an additional 45 minutes, followed by three PBS rinses.

Techniques: Staining, Expressing