Journal: The Journal of Biological Chemistry

Article Title: F2,6BP restores mitochondrial genome integrity in Huntington’s disease

doi: 10.1016/j.jbc.2026.111156

Figure Lengend Snippet: PFKFB3 and F2,6BP levels are low in HD patients impacting mitochondrial health. A , Western blot showing the relative levels of PFKFB3 in the cytosolic (CE), nuclear (NE) and mitochondrial (ME) extract of HEK293 cells. GAPDH: cytosolic loading control; HDAC2: nuclear loading control. COX4: mitochondrial loading control. B , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of Q-7 and Q-111 cells. COX4: mitochondrial loading control. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). C , Western blot showing the relative levels of PNKP and PFKFB3 in the mitochondrial extract of HD patients vs. age-matched control subjects’ frontal cortex. COX4: mitochondrial loading control. In the PFKFB3 immunoblot, upper band indicates a non-specific higher molecular weight cross-reactive band. Lower panel : Quantitation of the relative PFKFB3 levels after normalization with loading control COX4 (n = 3, ∗∗∗ p < 0.005). D , Benzonase-treated mitochondrial extracts from Q-7 cells were immunoprecipitated (IP’d) with anti-PFKFB3 antibody (PFKFB3 IP); or control IgG and tested for the presence of associated proteins (shown in the right) using specific Abs. Input: 10% of the extract used for IP. E-G , Q-7 ( E ) and Q-111 ( F ) cells were first stained with MitoTracker dye, which marks mitochondria in green ( left panels ), followed by fixation and staining for PFKFB3 using a secondary antibody conjugated to Alexa Fluor 568 ( middle panels ). The overlap of these signals appears yellow in merged images ( right panels ), confirming PFKFB3's mitochondrial localization. G , represents Q-111 cells after treatment with F2,6BP (50 μM), showing improved co-localization in the merged image suggesting that F2,6BP may help restore mitochondrial integrity and PFKFB3 levels in Q-111 cells. Nuclei are counterstained with DAPI. H , quantitation of the PFKFB3 level (n = 3, ∗∗ p < 0.01; ∗ p < 0.05). I , bar diagram showing the relative levels of F2,6BP in the mitochondrial extract of control vs. HD patients’ frontal cortex (n = 14, ∗ p < 0.05).

Article Snippet: The antibodies and chemicals used for immunostaining were as follows: Mouse Anti-HTT antibody-MW8 (Creative Biolab), Tetramethylrhodamine Methyl Ester Perchlorate (TMRM) from ThermoFischer scientific, USA, Thioflavin-T (Sigma-Aldrich), DAPI (Novus), NucRed dye (647) and MitoTracker from Invitrogem).

Techniques: Western Blot, Control, Quantitation Assay, Molecular Weight, Immunoprecipitation, Staining

Journal: The Journal of Biological Chemistry

Article Title: F2,6BP restores mitochondrial genome integrity in Huntington’s disease

doi: 10.1016/j.jbc.2026.111156

Figure Lengend Snippet: Exogenous F2,6BP alleviates pathogenic aggregates in HD cells. A , benzonase-treated mitochondrial extracts from Q-7, mock and F2,6BP-treated (100 μM, 72 h) Q-111 cells were immunoprecipitated (IP’d) with anti-PNKP antibody (PNKP IP); or control IgG and tested for the presence of associated proteins (shown in the right ) using specific Abs (n = 3). B , HTT aggregation levels in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP (100 μM; bottom ) were assessed with ThT dye, which fluoresces green , and nuclear localization was indicated by NucRed dye, providing a cyan signal in the merged images. Imaging was performed on the EVOS M5000 system at 200X magnification. ( Bottom ) Relative quantification of the mean fluorescence intensity (in arbitrary units) (∗ p < 0.05, between Q-7 and Q-111 cells and Q-111 cells + F2,6BP; ns = non-significant, p > 0.05, between Q-7 vs Q-111 (F2,6BP supplemented) cells. C , immunofluorescence micrographs show HTT expression and aggregation in Q-7 ( top ), Q-111 ( middle ) and Q-111 cells treated with F2,6BP ( bottom ). Cells were stained with Anti-HTT antibody MW8, visualized using a mouse secondary antibody conjugated to Alexa Fluor568, resulting in red fluorescence. The nuclei were counter stained with DAPI. Images were captured at 600X magnification using a SoRa super-resolution spinning disk confocal system with motorized FRAP/photobleaching.

Article Snippet: The antibodies and chemicals used for immunostaining were as follows: Mouse Anti-HTT antibody-MW8 (Creative Biolab), Tetramethylrhodamine Methyl Ester Perchlorate (TMRM) from ThermoFischer scientific, USA, Thioflavin-T (Sigma-Aldrich), DAPI (Novus), NucRed dye (647) and MitoTracker from Invitrogem).

Techniques: Immunoprecipitation, Control, Imaging, Quantitative Proteomics, Fluorescence, Immunofluorescence, Expressing, Staining

Journal: eLife

Article Title: Complete suspension culture of human induced pluripotent stem cells supplemented with suppressors of spontaneous differentiation

doi: 10.7554/eLife.89724

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , DAPI solution , FUJIFILM Wako Pure Chemical Corporation , Cat#340-07971 , .

Techniques: Recombinant, Suspension, Plasmid Preparation