Journal: Frontiers in Genetics

Article Title: Aberrant expression of the MID1 protein in neurons of Huntington’s disease brain

doi: 10.3389/fgene.2026.1753495

Figure Lengend Snippet: MID1 mRNA expression is aberrantly increased in HD-neurons. The MID1 expression was quantified in distinct cell populations isolated by MACS from the cortex of 15-week-old HdhQ150 knock-in mice (tg) and age-matched wild-type littermates (wt). The success of separation was quantified by qPCR analysis detecting GFAP as marker for astrocytes (A) , CD11b as marker for microglia (B) , and NeuN as neuronal marker (C) . (D) The marker proteins GFAP, NeuN, and CD11b were also detected on a Western blot of pooled samples from the tg animals (n = 7). MID1 was detected on the same blots. MID1 qPCR analysis revealed no statistically significant differences in (E) astrocytes ( p = 0.595) and (F) microglia ( p = 0.1966). (G) In neurons a significant upregulation of MID1 expression was detected (* p = 0.0104). MID1 levels were normalized to ACTB . Columns represent mean values ± SEM, p -values were calculated using an unpaired t-test with Welch’s correction (n wt = 7, n tg = 7).

Article Snippet: The Anti-ACSA-2 MicroBead Kit (130-097-678), the CD11b MicroBeads Kit (130-093-634), and the Neuron Isolation Kit (130-097-678) from Miltenyi Biotech were used for MACS.

Techniques: Expressing, Isolation, Knock-In, Marker, Western Blot

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Immunostaining of HMC3 cells with DAPI (blue) and IBA1 (red) after 24 hours of EV treatment. Scale bar: 100 μm; zoom scale bar: 50 μm (b-c). Percentage and surface area of activated HMC3 cells after 24 hours of EV treatment. Data are represented as mean ± SEM and median with interquartile range (n=8 images per group). P-values are represented in the figures. (d) Heatmap showing normalized signal intensity of cytokine expression in HMC3 cells treated with PD-EV- and HC-EV . (e) Western blot quantification of p16 INK4a and p21 WAF /CIP in HMC3 cells after 48 and 96 hours of EV treatment (n=3). (f-g) Semi-quantification of p16 INK 4a and p21 WAF /CIP expressed as percentage change: (Normalized signal (EV-treated))/ (Normalized signal (PBS control)-1) *100. Data are represented as mean ± SEM. P-value is represented in the figures.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Immunostaining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a-c) p16 INK4a, p21 WAF /CIP , and SERPINE1 mRNA relative expression in HMC3 cells after 48 and 96 hours of treatment with HC-EV, PD-EV, or PBS.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Expressing

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Schematic experimental setup of EV treatment on HMC3 cells followed by CM isolation and SH-SY5Y neurons treatment. (b,f) Immunostaining with DAPI (blue) and TUBB3 (red) of SH-SY5Y neurons after treatment with CM and EV - CM. Scale bar: 200 µm. (c,g) Neurite length quantification of SH-SY5Y neurons after 24 hours treatment with CM and EV - CM (n= 550-580 neurons per group). (d,h) Flow cytometry analysis of SH-SY5Y neurons exposed to CM and EV - CM for 24 hours. Annexin V FITC (x-axis) and propidium iodide (y-axis). Q1: dead cells (PI positive), Q2: late-stage apoptotic cells (Annexin V and PI positive), Q3: early-stage apoptotic cells (Annexin V positive), Q4: live cells (Annexin V and PI negative). (e,i) Bar graph showing percentage of cells in Q1 and Q2 stages after 24 hours treatment with CM and EV - CM. Results correspond to 3 replicates from at least three independent differentiations. (j) Neurite length and (k) apoptosis comparison of SH-SY5Y neurons treated with CM and EV - CM, respectively. Scatter plots show all data points, with median as a straight line and the IQR as an error bar. Bar plots show mean ± SEM. P-values are represented in the figures.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Isolation, Immunostaining, Flow Cytometry, Comparison