rabbit anti-rtn4  (FineTest Biotech Inc)

Journal: bioRxiv

Article Title: Leishmania exploits the macrophage endoplasmic reticulum-shaping protein CLIMP-63 to modulate mitochondrial biogenesis and bioenergetics

doi: 10.64898/2026.03.19.712868

Figure Lengend Snippet: BMM (treated with siRNA to either CLIMP-63 or RTN4 or treated with control siRNA, Ctr) were infected with L. donovani or L. amazonensis metacyclic promastigotes and at various time points post-phagocytosis parasite replication and PV size were assessed. (A) Quantification of L. donovani parasite burden in CLIMP-63 depleted BMM at 6, 24, 48, and 72 h post-infection. (B) Quantification of L. amazonensis parasite burden in CLIMP-63 depleted BMM at 6, 24, 48, and 72 h post-infection. (C) Quantification of PV size in CLIMP-63-depleted BMM infected with L. amazonensis at 48 and 72 h post-phagocytosis. (D) Quantification of L. donovani parasite burden in RTN4-depleted BMM at 6, 24, 48, and 72 h post-infection. (E) Quantification of L. amazonensis parasite burden in RTN4-depleted BMM at 6, 24, 48, and 72 h post-infection. (F) Quantification of PV size in RTN4-depleted BMM infected with L. amazonensis at 48 and 72 h post-phagocytosis. Data in (A, B, D, E) are presented as the means ± SEM of values of one representative experiment of three independent experiments. Data in (C, F) are presented as a violin plot with means ± standard deviations (SD) of values from three independent experiments for a total of 450 PVs. Statistics were calculated using one-way analyses of variance (ANOVA) with Sidak’s multiple comparison test with * P ≤ 0.05, *** P ≤ 0.001 and **** P ≤ 0.0001 significance. Blots showing the efficacy the siRNA-mediated CLIMP-63 and RTN4 knockdowns are shown in S3 Fig and S4 Fig, respectively.

Article Snippet: The mouse anti-CLIMP-63 monoclonal antibody (sc-393544) was from Santa Cruz Biotechnology, the rabbit anti-RTN4 polyclonal antibody (ab186735) and the rat anti-BrdU (ab6326) were from Abcam, the rabbit polyclonal antibody RTN4 (10950-1-AP) was from Proteintech, the mouse anti-phosphoglycan (Galβ1,4Manα1-PO4) CA7AE monoclonal antibody ( ) was from Cedarlane, the rabbit anti-β-actin polyclonal antibody was from Cell Signalling, the rabbit anti-Tom20 polyclonal antibody (EPR15581-54) was from Abcam, and the rat anti-LAMP-1 monoclonal antibody 1D4B developed by J.T.

Techniques: Control, Infection, Comparison

Journal: bioRxiv

Article Title: TBC1D24 regulates mitochondria and endoplasmic reticulum-mitochondria contact sites

doi: 10.1101/2024.09.19.613961

Figure Lengend Snippet: (A-B) Representative images showing the localisation of Flag-TBC1D24 (Flag, Green) in transfected primary fibroblasts, along with mitochondria (TOM20, Magenta)(A) or ER tubules (RTN4, Magenta)(B). (C-D) TBC1D24 mutations affect the structure of ER tubules. (C) Representative images ER tubules (RTN4, Red) and nuclei (Dapi, Blue) in control and patient fibroblasts. Scale bars 10 µm (D) Quantification of ER tubule phenotypes from images in (C). Cells with ER tubules similar to the patient fibroblast shown in (C) were considered as altered. Each point represents an individual experiment. Bars show the average ± SD. *** P<0.001. One-way ANOVA. (E-F) TBC1D24 mutations affect the structure of ER sheets. (E) Representative images ER tubules (CLIMP63, Red) and nuclei (Dapi, Blue) in control and patient fibroblasts. Scale bars 10 µm (F) Quantification of ER sheets phenotypes from images in (E). Cells with ER tubules similar to the patient fibroblasts shown in (E) were considered as altered. Each point represents an individual experiment. Bars show the average ± SD. * p<0.05, *** P<0.001. One-way ANOVA.

Article Snippet: The cells were then incubated for 1 hour at room temperature in blocking buffer with the following primary antibodies: TOM20 (Rb, Abcam, ab186735, 1:250), CLIM63 (Mo, ENZO, ENZ-ABS669, 1: 200), RTN4 (Rb, Bio-Rad, AHP1799, 1:200), FLAG M2 (Mo, Sigma-Aldrich, F1804, 1:200).

Techniques: Transfection, Control