rabbit anti-rab5c  (ABclonal Biotechnology)

Journal: Genome Biology

Article Title: Surfaceome CRISPR screen identifies OLFML3 as a rhinovirus-inducible IFN antagonist

doi: 10.1186/s13059-021-02513-w

Figure Lengend Snippet: Validation of the effects of ICAM-1, RAB5C, and OLFML3 on RV infection. a , b Time-dependent viral replication of RV-B14 in mock and knockout cells, as determined by viral loads in medium supernatant ( a ) and cell lysate ( b ). Cells are infected with RV-B14 at an MOI of 2. Viral RNA in cell lysates ( b ) is normalized to RPLP0 expression. Significant difference between test groups and non-targeting sgRNA group is determined using two-way ANOVA with Dunnett’s multiple comparisons test. c , d Rescued susceptibility of knockout cells to the infection of RV-B14 ( c ) and RV-A16 ( d ) by overexpression of RAB5C and OLFML3 respectively. e , f Rescued replication of RV-B14 ( e ) and RV-A16 ( f ) in knockout cells by overexpression of RAB5C and OLFML3 respectively, as determined by viral loads in medium supernatant. g , h Rescued replication of RV-B14 ( g ) and RV-A16 ( h ) in knockout cells by overexpression of RAB5C and OLFML3 respectively, as determined by viral loads in cell lysates. Viral RNA in cell lysates is normalized to RPLP0 expression. i Representative images of CPEs induced by clinical RV strain. Scale bar, 100 μm. j Cell viability of mock and knockout cells upon challenge of clinically isolated RV strain. k Phylogenetic analyses of clinical RV strain using MEGA X , with VP4 gene as the reference. l , m The effects of gene knockout on the replication of clinical RV strain, as determined by RT-qPCR quantification of viral loads in medium supernatant ( l ) or cell lysates ( m ). For m , viral RNA in cell lysates is normalized to RPLP0 expression. For c – j and l , m , analyses are performed at 24 h post infection of RV at an MOI of 2. Significant difference between test groups and non-targeting sgRNA group is determined using two-tailed Student’s t test and the P values are shown. Significant difference between knockout and overexpression rescue groups is determined using two-tailed Student’s t test and the P values are shown above the lines

Article Snippet: The following primary and secondary antibodies were used in WB: anti-ICAM1 rabbit antibody (Cell Signaling Technology, Cat. No. 4915S, Danvers, USA), anti-RAB5C rabbit antibody (Thermo, Cat. No. PA551932), anti-OLFML3 rabbit antibody (Thermo, Cat. No. PA531581), HRP-conjugated anti-rabbit IgG (CST, Cat. No. 7074S).

Techniques: Biomarker Discovery, Infection, Knock-Out, Expressing, Over Expression, Isolation, Gene Knockout, Quantitative RT-PCR, Two Tailed Test

Journal: Genome Biology

Article Title: Surfaceome CRISPR screen identifies OLFML3 as a rhinovirus-inducible IFN antagonist

doi: 10.1186/s13059-021-02513-w

Figure Lengend Snippet: Dissection of the functions of RAB5C and OLFML3 in RV infection. a , b Viral loads in medium supernatant ( a ) and cell lysates ( b ) at 24 h after transfection of RV-A16 genome RNA. c , d Viral RNA in cell lysates of mock and knockout cells at 1, 3, and 6 h after infection with RV-B14 ( c ) and RV-A16 ( d ) at an MOI of 20 in the presence of 2 mM GuHCl. For a , b , significant difference between mock and test groups is determined using two-tailed Student’s t test. For c , d , significant difference between mock and RAB5C groups is determined using two-tailed Student’s t test. e Volcano plot showing differentially expressed genes (DEGs). RV infection-induced gene upregulation and downregulation are first calculated and the differentially upregulated or downregulated genes in mock and knockout cells are defined as DEGs. Cells are harvested and analyzed at 24 h after infection of RV-B14 at an MOI of 2. f GO analyses of biological processes of DEGs identified in E. g , h Heat map showing ISG expression in mock and OLFML3 −/− cells at 24 h post infection of RV-B14 ( g ) and RV-A16 ( h ) at an MOI of 2. These results are derived from RT-qPCR quantification. Gene expression is normalized to RPLP0

Article Snippet: The following primary and secondary antibodies were used in WB: anti-ICAM1 rabbit antibody (Cell Signaling Technology, Cat. No. 4915S, Danvers, USA), anti-RAB5C rabbit antibody (Thermo, Cat. No. PA551932), anti-OLFML3 rabbit antibody (Thermo, Cat. No. PA531581), HRP-conjugated anti-rabbit IgG (CST, Cat. No. 7074S).

Techniques: Dissection, Infection, Transfection, Knock-Out, Two Tailed Test, Expressing, Derivative Assay, Quantitative RT-PCR, Gene Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sorcin regulates alveolarization and airway tissue remodeling during lung morphogenesis

doi: 10.1007/s00018-025-05870-y

Figure Lengend Snippet: Altered protein expression and EGFR signaling in Sorcin-deficient lung development A ) Confocal microscopy of 3-week-old lung tissue from WT and Sri⁻ / ⁻ mice. Sri⁻ / ⁻ lung tissue shows reduced SP-B (red) expression compared to WT. Scale bars, 20 μm B ) Western blot analysis of lung lysates from 3-week-old WT and Sri⁻ / ⁻ mice. Sri⁻ / ⁻ mice exhibit decreased EGFR, PANRAS and RAB5C protein levels compared to WT. Densitometry quantification is shown (mean ± SEM). ** p < 0.01 (Student’s t-test), *** p < 0.001 (determined by Student’s t-test) C-E ) Confocal microscopy of 3-week-old lung sections: WT mice show epithelial localization of EGFR (green) (C), RAB5C (green) (D) and PANRAS (red) protein (E). In Sri⁻ / ⁻ mice, these proteins are significantly reduced. Scale bars, 20 μm

Article Snippet: Then, sections were blocked in 5% goat serum for 1 h and incubated overnight at 4 °C with primary antibody rabbit polyclonal EPCAM/CD326 (1:100 in 1%BSA-PBS solution) (Proteintech, #21050-1-AP), rabbit polyclonal SFTPB (1:100 in 1%BSA-PBS solution) (Thermo Fisher Scientific, # PA542000 ), rabbit polyclonal E-cadherin (1:100 in 1%BSA-PBS solution) (Proteintech, #20874-1-AP), mouse monoclonal E-cadherin (1:100 in 1%BSA-PBS solution) (BD Bioscience, #610181), mouse monoclonal α-Smooth Muscle Actin (1:100 in 1%BSA-PBS solution) (Sigma-Aldrich, #A5228), rabbit polyclonal EGFR (1:100 in 1%BSA-PBS solution) (Proteintech, #30847-1-AP) rabbit polyclonal RAB5C (1:100 in 1%BSA-PBS solution) (Thermo Fisher Scientific, #PA5101828), mouse monoclonal PANRAS (Ab3) (1:100 in 1%BSA-PBS solution) (Sigma-Aldrich, #OP40) followed by incubation with Alexa fluor 488 (rabbit)-conjugated secondary antibodies (1:500 in 1%BSA-PBS solution) (Thermo Fisher Scientific).

Techniques: Expressing, Confocal Microscopy, Western Blot

Journal: Cell Reports Medicine

Article Title: A genome-wide association study identified PRKCB as a causal gene and therapeutic target for Mycobacterium avium complex disease

doi: 10.1016/j.xcrm.2024.101923

Figure Lengend Snippet:

Article Snippet: Rabbit anti -RAB5C , ABclonal , Cat#A7342; RRID: AB_2767879.

Techniques: Virus, Recombinant, Modification, Magnetic Beads, Protease Inhibitor, Western Blot, Reporter Assay, Gene Expression, Generated, Plasmid Preparation, Software

Journal: EBioMedicine

Article Title: Paricalcitol accelerates BACE1 lysosomal degradation and inhibits calpain-1 dependent neuronal loss in APP/PS1 transgenic mice

doi: 10.1016/j.ebiom.2019.07.014

Figure Lengend Snippet: PAL treatment decreases BACE1 expression in APP/PS1 mice. (a) Sections from APP/PS1 mouse brians were co-stained with VDR (green) and NeuN (red); the merged image from cortex shows the predominant localization of VDR in NeuN-positive neurons, and the large arrows and small arrows show the localization of VDR in the epithelium and glial cells, respectively. (b-c) Inhibition of total and nuclear SREBP2 were associated with VDR activation in the cortex of APP/PS1 mice. n = 8. (d) PAL treatment dramatically suppressed the expression of BACE1 but caused no significant differences in the protein levels of APP, ADAM10 or PS1 in APP/PS1 mouse brains. n = 8. (e) Immunoblotting showed that the protein levels of sAPPβ and C99 are decreased, but the protein levels of sAPPα and C83 are unchanged in APP/PS1 mouse brains after PAL treatment. n = 8. (f) APP and BACE1 mRNA expression levels were not significantly different in APP/PS1 mouse brains after PAL treatment. n = 6. (g) PAL treatment induced a marked upregulation of LRP1 without altering the expression of APOE, RAGE, IDE and NEP in APP/PS1 mouse brains. n = 8. (h) PAL treatment increased the immunointensity of LAMP1 in CA3 region. Three sections/brain, n = 6. (i) The lysosomal markers (LAMP1 and LAMP2) were increased in APP/PS1 mouse cortex after PAL treatment. n = 6. (j) Co-localizations of BACE1 and LAMP1 were increased in CA3 region after PAL treatment. The white arrows show BACE1 is not co-localized with LAMP1. Three sections/brain, n = 5. (k) The expression levels of rab5c (early endosomal marker) and rab7a (late endosomal marker) were significantly reduced in APP/PS1 mouse brains after PAL treatment. n = 8. * p < .05, ** p < .01 (Student's- t -test). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Equal proteins were subjected to SDS/PAGE to separate, the proteins were transferred to PVDF membranes and incubated with mouse anti-VDR (Santa Cruz, sc-13,133, 1: 500), rabbit anti-BACE1 (Abcam, ab183612, 1: 1000), rabbit anti-APP695 (Cell Signaling Technology, 2452, 1: 1000), rabbit anti-C-APP (Sigma, SAB4200535, 1:1000), rabbit anti-PS1 (Cell Signaling Technology, 5643, 1: 1000), rabbit anti-AMAD 10 (Cell Signaling Technology, 14,194, 1: 1000), mouse anti-soluble amyloid precursor α (sAPPα; Immuno-Biological Laboratories, 11,088, 1: 500), mouse anti-soluble amyloid precursor β (sAPPβ; Immuno-Biological Laboratories, 10,321, 1: 500), rabbit anti-advanced glycation end products (RAGE; Sigma, SAB2105049, 1: 1000), goat anti-apolipoprotein (APOE; Santa Cruz, sc-6384, 1: 500), mouse anti-LRP1 (Santa Cruz, sc-57,353, 1: 400), rabbit anti-LRP1 (Abcam, ab92544, 1: 8000), mouse anti-insulin-degrading enzyme (IDE; Santa Cruz, sc-514,458, 1:500), mouse anti-neprilysin (NEP; Santa Cruz, sc-46,656, 1: 500), mouse anti-HSP70 (Themo Fisher Scientific, MA3–008, 1: 2000), rabbit anti-Rab5c (Themo Fisher Scientific, PA5–36606, 1: 500), goat anti-Rab7a (Biorbyt, orb180471, 1: 1000), rabbit anti-postsynaptic density proteins 95 (PSD95; Cell Signaling Technology, 3409, 1: 2000), mouse anti-synaptophysin (SYP; Santa Cruz, sc-365,488, 1:1000), mouse anti-OGG1 (Santa Cruz, sc-376,935, 1: 500), mouse anti-MTH1 (Santa Cruz, sc-271,082, 1: 500), mouse anti-MUTYH (Santa Cruz, sc-374,571, 1: 500), rabbit anti-LAMP1 (Abcam, ab24170, 1: 1000), rabbit anti-LAMP2 (Abcam, ab18528, 1: 1000), rabbit anti-NeuN (Cell Signaling Technology, 12,943 s, 1: 2000), rabbit anti-IL-1β (Santa Cruz, sc-7884, 1: 500) or mouse anti-TNFα (Santa Cruz, sc-52,746, 1: 500) overnight at 4 °C.

Techniques: Expressing, Staining, Inhibition, Activation Assay, Western Blot, Marker