antibody against lactb2  (Proteintech)

Journal: bioRxiv

Article Title: Carbohydrate adaptation drives liver-brain axis maturation

doi: 10.1101/2025.11.05.685548

Figure Lengend Snippet: a. Top-ranked functional groups of differential genes significantly associated with developmental progression. b. Transmission electron microscopy (TEM) images of hepatic mitochondria across postnatal stages. Scale bar, 0.2 um. c. Comparison of ETC complex activity measured as oxygen consumption rate in isolated liver mitochondria across postnatal stages ( N = 4-6 per group). Data are means ± SEM and analyzed by two-tailed unpaired t-test, * P < 0.05, **** P < 0.0001. d. Waterfall plot of nuclear-encoded mitochondrial genes involved in mtDNA gene regulation and ETC function, ranked by their correlation with developmental progression ( P < 0.05). e. Quantitative RT-PCR of Lactb2 expression in developing livers (mean ± SEM, N = 4 per stage). Two-tailed unpaired t-test compared of each stage to P0. * P < 0.05, ** P < 0.01. f. Immunoblot analysis of LACTB2, ETC complex proteins and mitochondrial VDAC in developing livers with vinculin as loading control. g. Immunohistochemical (IHC) staining of LACTB2 in major mouse organs using mouse tissue microarrays. Scale bar, 50 µm. h. scRNA-seq scatter plot showing Lactb2 expression levels across distinct hepatic cell populations during postnatal development.

Article Snippet: After fixation with fresh warm 4% paraformaldehyde in PBS for 15 minutes at room temperature, cells were permeabilized in 0.1% (v/v) Triton X-100 in PBS for 10 minutes, pre-blocked with filtered 1% BSA (w/v) in PBS for 1 hour and then incubated with primary antibody against LACTB2 (1:200, Proteintech, 16783-1-AP), and HPS60 (1:100, ThermoFisher, MA3-012) diluted in the blocking buffer for 1 hour at room temperature.

Techniques: Functional Assay, Transmission Assay, Electron Microscopy, Comparison, Activity Assay, Isolation, Two Tailed Test, Quantitative RT-PCR, Expressing, Western Blot, Control, Immunohistochemical staining, Immunohistochemistry

Journal: bioRxiv

Article Title: Carbohydrate adaptation drives liver-brain axis maturation

doi: 10.1101/2025.11.05.685548

Figure Lengend Snippet: a. Northern blot analysis of mt-RNAs in the indicated isogenic H2.35 hepatocytes. Nuclear 18S rRNA is used as a loading control. b. Blue-Native PAGE analysis of ETC supercomplexes in mitochondria isolated from control and LACTB2-depleted H2.35 hepatocytes. c. ETC complex activity assessed by oxygen consumption rates in the presence of complex-specific substrates in the indicated H2.35 hepatocytes. Results are means ± SEM ( N = 6) and p values are calculated by two-tailed unpaired t-test, * P < 0.05, ** P < 0.0, **** P < 0.0001, ns, not significant. d. Immunoblot analysis of ETC complex proteins in LACTB2-depleted hepatocytes with or without LACTB2 re-expression. The cells were treated with tigecycline at 50 uM for 72h, followed by drug withdraw for the indicated hours. “-” indicates DMSO treatment. e. Survival of WT, Het and KO male and female mice. N numbers are indicated for each group; P value by log-rank Mantel-Cox test. f. Body weight gain up to 6 weeks postpartum. Data are mean ± SD ( N = 4-6 per group) and *** P < 0.001 by mixed-effects model. g. TEM of WT and KO male livers at P35 ( N = 3 per group). Scale bar: 0.5 µm. h. ETC complex activity measured by OCR in isolated liver mitochondria from WT, Het and KO males. Results are mean ± SEM ( N = 3-4 mice per group) and analyzed by two-tailed unpaired t-test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, n.s. not significant. i. Relative abundance of mitochondrial rRNAs and mRNAs in WT, Het and KO male livers at P35 as assessed by strand-specific RNA-seq ( N = 4 per group). Results are mean ± SD and p values are calculated by two-tailed unpaired t-test, * P < 0.05, ** P < 0.01. j. Blue-Native PAGE analysis of ETC supercomplexes using mitochondria isolated from WT and KO male livers at P35. k. Immunoblot analysis of ISR markers in WT and KO male mouse livers at P35 ( N = 3 per group). l. Survival of KO male mice treated with AAV8-LACTB2, AAV8-NDI1 and ISRIB. N numbers are indicated for each group. P values by log-rank Mantel-Cox test: P < 0.0001 for AAV8-LACTB2 and AAV8-NDI1; P = 0.0022 for ISRIB.

Article Snippet: After fixation with fresh warm 4% paraformaldehyde in PBS for 15 minutes at room temperature, cells were permeabilized in 0.1% (v/v) Triton X-100 in PBS for 10 minutes, pre-blocked with filtered 1% BSA (w/v) in PBS for 1 hour and then incubated with primary antibody against LACTB2 (1:200, Proteintech, 16783-1-AP), and HPS60 (1:100, ThermoFisher, MA3-012) diluted in the blocking buffer for 1 hour at room temperature.

Techniques: Northern Blot, Control, Blue Native PAGE, Isolation, Activity Assay, Two Tailed Test, Western Blot, Expressing, RNA Sequencing