Journal: CNS Neuroscience & Therapeutics

Article Title: Rapamycin Reduces Amyloid‐β Plaques and Improves Behavioral Performance in a Sex‐Dependent Manner in Mouse Models of Amyloidosis

doi: 10.1002/cns.70807

Figure Lengend Snippet: Reduced amyloid plaque‐associated toxicity in rapamycin‐treated 5xFAD mice. (A) Confocal images showing CD68 (green) and Iba1 (red) staining in the brains of vehicle‐ or rapamycin‐treated female and male 5xFAD mice. Scale bar, 500 μm. (B) Quantification of the CD68 + /Iba1 + area ratio. n = 5 or 6 mice per group. Female: T (10) = 4.063, p = 0.0023; male: T (8) = 1.657, p = 0.1361, unpaired t ‐test. (C) Confocal images showing LAMP1 (green) staining in the cells. Scale bar, 500 μm. (D) Quantification of the LAMP1 + area. n = 6 mice per group. Female: T (10) = 3.718, p = 0.0040; male: T (10) = 0.1617, p = 0.8748, unpaired t ‐test. (E) Confocal images showing glial fibrillary acidic protein (GFAP; red) expression. Scale bar, 500 μm. (F) Quantification of the GFAP + area. n = 6 mice per group. Female: T (10) = 2.481, p = 0.0325; male: T (10) = 0.08114, p = 0.9369, unpaired t ‐test. Data are mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: LAMP1 , DSHB , Cat#1D4B; RRID: AB_528127.

Techniques: Staining, Expressing

Journal: bioRxiv

Article Title: Small-molecule CBLB inhibitor abolishes EGFR ubiquitination, reduces receptor endocytosis and diminishes cell motility signaling

doi: 10.64898/2026.02.24.707836

Figure Lengend Snippet: Cells were pre-incubated with DMSO or 100 nM NX-1013 and then incubated with 4 ng/ml EGF-Rh for 10 or 60 min at 37°C in presence or absence of the inhibitor, fixed, permeabilized and labeled with EEA1 ( A ) or LAMP1 ( B ) antibodies. Imaging was through the 640 nm ( green , EEA1/LAMP1), 561 nm ( red , EGF-Rh) and 405 nm ( blue , nuclei) channels. Maximum intensity projections of z-stacks are presented. Fluorescence intensity scales are identical for all images. White arrows mark examples of EGF-Rh puncta colocalized with EEA1 or LAMP1. Arrowheads point to the plasma membrane. Scale bars, 10 μm. ( C ) Quantifications of the fraction of EGF-Rh co-localized with EEA1 or LAMP1 of total cell-associated EGF-Rh in images exemplified in ( A ) and ( B ). Bar graph represents mean values (± SEM; n=4-6 FOVs) of the fraction of EGF-Rh co-localized with EEA1 or LAMP1. Unpaired two-tailed Student T-test between vehicle and NX-1013-treated cells for the same incubation times was used to calculate p values. *p<0.05 (p=0.019); **p<0.01 (p=0.0095); ****p<0.0001; ns, not significant (p>0.05).

Article Snippet: The antibody to CHC (ab21679) and rabbit EEA1 (ab2900) were purchased from Abcam, to Grb2 (Rb PA5-27151) from Invitrogen and to LAMP1 (H4A3) from DSHB.

Techniques: Incubation, Labeling, Imaging, Fluorescence, Clinical Proteomics, Membrane, Two Tailed Test

Journal: The Journal of Cell Biology

Article Title: Rsp5/NEDD4 and ESCRT regulate TDP-43 toxicity and turnover via an endolysosomal clearance mechanism

doi: 10.1083/jcb.202212064

Figure Lengend Snippet: Mild TDP-43 overexpression causes accumulation of giant Rab7-positive vesicles. (A) HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells were grown in normal conditions before being fixed and immunostained for Rab7. N = 3; scale bar, 10 µm. (B) Quantification of the frequency of giant endosomes in HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells. **, P < 0.01; ***, P < 0.001 by one-way analysis of variance with Tukey’s post hoc test; N = 3. (C) Quantification of the average size of giant endosomes in HEK293A WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cells. ***, P < 0.001 by one-way analysis of variance with Tukey’s post hoc test; N = 3. (D) Distribution of giant Rab7+ve vesicle diameters from WT, HEK-TDP-43-GFP, and HEK-TDP-35-GFP cell lines. N = 3. (E) HEK-TDP-43-GFP (pseudocolored white) cells were grown in normal conditions before being fixed and immunostained for Rab5, LC3B, or LAMP1 (pseudocolored green) and then successively immunostained for Rab7 (pseudocolored red). N = 3; scale bar, 10 µm. (F) HEK-TDP-35-GFP (pseudocolored white) cells were grown in normal conditions before being fixed and immunostained for Rab5, LC3B, or LAMP1 (pseudocolored green) and then successively immunostained for Rab7 (pseudocolored red). N = 3; scale bar, 10 µm in main panels; 2 µm in insets. Error bars in all graphical panels represent standard error of the mean.

Article Snippet: Primary antibodies used for the immunofluorescence were Rab7 (ab137029; Abcam, dilution: 1:200), CD63 (H5C6; Developmental Studies Hybridoma Bank, dilution 1:100), Rab5 (ab109534; Abcam, dilution: 1:200), LC3B (3868S; Cell Signaling, dilution 1:500), LAMP1 (21997-1-AP; Proteintech, dilution 1:200), and TDP-43 (10782-2-AP; Proteintech; dilution 1:200).

Techniques: Over Expression