Journal: Journal of ethnopharmacology

Article Title: Extract of Nanhaia speciosa J. Compton & Schrire alleviates LPS-induced acute lung injury via the NF-κB/Nrf2/AQPs pathway.

doi: 10.1016/j.jep.2024.118831

Figure Lengend Snippet: Fig. 7. The expression levels of Keap1, Nrf2, HO-1 and NQO1 in A549 cells were detected by Western blotting. (A) Keap1, (B) Cytoplasm Nrf2, (C) Nucleus Nrf2, (D) NQO1, and (E) HO-1. (F) The expression of Nrf2 was detected by immunofluorescence, and the scale bar distance in the figure is 20 μm. Data are means ± SD, n = 3. *p < 0.05 compared with control group the difference was statistically significant.

Article Snippet: Antibodies against HO-1 (10701- 1-AP), Nrf2 (80593-1-RR), NQO1 (67240-1-Ig), TRAF2 (67315-1-Ig), NF-κB p65 (80979-1-RR), β-actin (66009-1-Ig), and Lamin B1 (66095-1- Ig) were obtained from Proteintech (Wuhan, China).

Techniques: Expressing, Western Blot, Immunofluorescence, Control

Journal: PLoS Pathogens

Article Title: Human Cytomegalovirus miR-UL148D Facilitates Latent Viral Infection by Targeting Host Cell Immediate Early Response Gene 5

doi: 10.1371/journal.ppat.1006007

Figure Lengend Snippet: (A) Kasumi-3 and CD34 + HPCs were efficiently infected with the NR-1 strain of HCMV. Kasumi-3 and HPCs were either mock-infected or infected with a GFP-expressing NR-1 strain at the indicated multiplicities of infection (MOIs). Two days later, the cells were analyzed for GFP expression by flow cytometry. An MOI of 5 was used for the following experiment. (B) The maintenance of the NR-1 genome, the suppression of viral IE1 and the presence of latency-associated UL138 over a 10-day time course. DNA and total RNA were isolated from Kasumi-3 cells and HPCs at various time points after infection. Viral genomic DNA was assayed by PCR, and RNA molecules encoding IE1 and UL138 were assayed by RT-PCR. In both cases, gel electrophoresis was used to detect the products of the reactions. (C) Reactivation of NR-1 virus in infected Kasumi-3 cells and HPCs. Kasumi-3 cells and HPCs were latently infected along a 10-day time course. Then, a subset of each cell population was cultured for an additional 2 days under conditions favoring lytic reactivation: Kasumi-3 cells were exposed to TPA, while HPCs were grown in reactivation medium. Following this, total RNA was extracted from the cells, and the ratio of IE1 to UL138 cDNA expression was assessed by qRT-PCR in triplicate. (D) Release of infectious progeny virions in latently infected Kasumi-3 cells and HPCs following reactivation treatment. Latently infected or mock-infected Kasumi-3 cells and HPCs were cultured under conditions favoring lytic reactivation (described above) or control conditions for 6 days, after which the cells were washed with PBS and co-cultured with HFFs for 2 days. Then, the Kasumi-3 cells were removed from the co-cultures, and the HFFs were washed with PBS and cultured for an additional 5 days for fluorescence microscopy analysis of GFP-positive plaques. (E) miR-UL148D showed robust accumulation during the establishment of experimental HCMV latency in Kasumi-3 cells. In total, 20,000 infected cells were harvested for the isolation of total RNA and DNA at each indicted time point along the 10-day time course. Viral DNA was first quantified by qPCR, and then absolute viral genomes copies were calculated by generating a standard curve. HCMV miRNAs were then assayed with a HCMV miRNA probe kit, and their levels were calculated using a standard curve. The HCMV miRNA level per virus was calculated by dividing the amount of each HCMV miRNA by the virus copy number. (F) miR-UL148D accumulated in HPCs latently infected with NR-1. HCMV miRNA levels in NR-1-infected HPCs were determined as described above. Values are shown as the mean ± SEM (n = 3). **, P<0.01.

Article Snippet: Anti-HCMV IgG and IgM antibodies in plasma were detected by ELISA using an HCMV IgG/IgM kit (DIA PRO Diagnostic Bioprobes, Milano, Italy) according to the manufacturer’s instructions.

Techniques: Infection, Expressing, Flow Cytometry, Isolation, Reverse Transcription Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Virus, Cell Culture, Quantitative RT-PCR, Control, Fluorescence, Microscopy

Journal: PLoS Pathogens

Article Title: Human Cytomegalovirus miR-UL148D Facilitates Latent Viral Infection by Targeting Host Cell Immediate Early Response Gene 5

doi: 10.1371/journal.ppat.1006007

Figure Lengend Snippet: (A, C) HCMV genome copies in Kasumi-3 cells (A) and CD34 + HPCs (C) infected with NR-1 or NR-1ΔmiR-UL148D. Total DNA was isolated from the infected cells at various time points after infection, and viral DNA was quantified by qPCR and normalized to cellular GAPDH. (B, D) Representative transcript levels from each class of viral genes in Kasumi-3 cells (B) and HPCs (D) infected with NR-1 or NR-1ΔmiR-UL148D. IE1 (immediately early), UL54 (early lytic transcript), and UL99 (late lytic transcript). Total RNA was isolated from infected cells and assayed by RT-qPCR. Samples were assayed in triplicate, and GAPDH level was used for normalization. (E, F) Restoring miR-UL148D expression via transfection with the miR-UL148D agomir reduced HCMV genome copies (E) and IE1 (F) expression in NR-1ΔmiR-UL148D-infected Kasumi-3 cells. The miR-UL148D agomir was added 24 hours before viral infection, and the culture media was replaced everyday with the addition of fresh agomir. DNA and total RNA were isolated from the Kasumi-3 cells at various time points after infection and quantified by qPCR and RT-qPCR, respectively. Samples were assayed in triplicate, and GAPDH level was used for normalization. Results derived from NR-1-infected Kasumi-3 cells are shown as a control. (G) Both NR-1ΔmiR-UL148D- and NR-1-infected Kasumi-3 cells produced infectious progeny. Infected Kasumi-3 cells harvested 10 days post-infection were stained with a monoclonal antibody against IE1 (clone 1B12, shown in red). GFP (green) and DAPI (blue) served as markers for lytic infection and nuclei, respectively. Infected Kasumi-3 cells were also co-cultured with HFFs. Viral plaque formation in the HFFs (shown by a GFP-positive status) was visualized by fluorescence microscopy. Images were collected using a 40x objective, and representative fields are shown for each infection. Values are shown as the mean ± SEM (n = 3). *, P<0.05. **, P<0.01.

Article Snippet: Anti-HCMV IgG and IgM antibodies in plasma were detected by ELISA using an HCMV IgG/IgM kit (DIA PRO Diagnostic Bioprobes, Milano, Italy) according to the manufacturer’s instructions.

Techniques: Infection, Isolation, Quantitative RT-PCR, Expressing, Transfection, Derivative Assay, Control, Produced, Staining, Cell Culture, Fluorescence, Microscopy

Journal: PLoS Pathogens

Article Title: Human Cytomegalovirus miR-UL148D Facilitates Latent Viral Infection by Targeting Host Cell Immediate Early Response Gene 5

doi: 10.1371/journal.ppat.1006007

Figure Lengend Snippet: (A) Schematic of miR-UL148D-binding sites on IER5, with the two seed-recognizing sites marked in red. The predicted free energy of each hybrid is indicated. (B) Downregulation of IER5 and upregulation of CDC25B protein levels in Kasumi-3 cells induced by miR-UL148D mimics. The protein levels of IER5 and CDC25B in Kasumi-3 cells 48 hours after transfection with miR-UL148D mimics were assayed with western blotting. GAPDH served as an internal control. (C) Upper panel: construction of luciferase reporter that expresses the IER5 3’UTR with or without mutation of miR-UL148D-binding sites. Mutations of the seed-recognizing sites in the IER5 3’UTR are marked in red. Lower panel: firefly luciferase activity in 293T cells that were co-transfected with various luciferase reporter constructs for the IER5 3’UTR and miR-UL148D, normalized to β-gal activity. (D) Upper panel: Construction of an expression vector harboring the ORF and 3’UTR of IER5 with or without mutation of the two miR-UL148D-binding sites. The mutations in the binding sites in the IER5 3’UTR are marked in red. Lower panel: Expression levels of IER5 and CDC25B in Kasumi-3 cells that were co-transfected with wild-type IER5 (IER5 WT) or mutated IER5 (IER5 Mut) and miR-UL148D or a scrambled oligonucleotide. Protein levels were detected by Western blot analysis after a 72-h transfection. The efficiency of transfection was monitored by assaying β-gal activity. (E) Schematic representing the hypothesis for how miR-UL148D facilitates HCMV latency. Values are shown as the mean ± SEM (n = 3). **, P<0.01.

Article Snippet: Anti-HCMV IgG and IgM antibodies in plasma were detected by ELISA using an HCMV IgG/IgM kit (DIA PRO Diagnostic Bioprobes, Milano, Italy) according to the manufacturer’s instructions.

Techniques: Binding Assay, Transfection, Western Blot, Control, Luciferase, Mutagenesis, Activity Assay, Construct, Expressing, Plasmid Preparation

Journal: Scientific Reports

Article Title: Caveolin-1α regulates primary cilium length by controlling RhoA GTPase activity

doi: 10.1038/s41598-018-38020-5

Figure Lengend Snippet: Cav1α controls cilium length via ROCK and Dia1. (A , B) Control and Cav1α KO cells were untreated (−) or treated with 10 µM Y27632 for 24 h and then stained for acetylated tubulin, F-actin and γ-tubulin ( A ). The scatter-plot represents the total length of cilia measured in µm; more than 700 cells were analyzed for each condition ( B ). (C) Control and DIA1 KO cells were analyzed by immunoblotting for DIA1. GAPDH was used as a loading control. (D , E) Control and DIA1 KO cells were grown for 5 days and stained for acetylated tubulin, F-actin, γ-tubulin and nuclei ( D ). The scatter-plot represents ciliary lengths measured in µm; more than 700 cells were analyzed in control and DIA1 KO cells ( E ). (F–H) Control and Cav1α KO cells were transiently transfected with the indicated constructs, then fixed and stained after 72 h for F-actin and acetylated tubulin ( F ). The scatter-plots represent the ciliary lengths measured in µm; more than 100 cells were analyzed for each condition ( G , H ). Scale bars, 5 µm. Data in B, E, G and H were pooled from at least three independent experiments and are represented as the mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, non-significant.

Article Snippet: The sources of the antibodies to the different markers were as follows: Cav1 (mouse mAb IgG1, used at 1/2,000 for immunoblot analysis; 610406; and the rabbit polyclonal antibody used at 1/200 for immunofluorescence analysis; 610059) and DIA1 (mouse mAb IgG1, used at 1/1000 for immunoblotting; clone51/mDia1; 610484), were from BD Transduction Laboratories. γ-tubulin (mouse mAb IgG1, used at 1/500 for immunofluorescence analysis; clone GTU-88, T3559) and acetylated tubulin (mouse mAb IgG2b, used at 1/500 for immunofluorescence analysis; clone 6-11B-1; T7451) were obtained from Sigma-Aldrich.

Techniques: Staining, Western Blot, Transfection, Construct

Journal: Scientific Reports

Article Title: Caveolin-1α regulates primary cilium length by controlling RhoA GTPase activity

doi: 10.1038/s41598-018-38020-5

Figure Lengend Snippet: Model of Cav1α function in the regulation of primary cilia length. In control cells (left), Cav1α localizes in non-caveolar domains at the apical membrane of MDCK cells, where it positively regulates RhoA activity. Active RhoA subsequently promotes apical actin polymerization through its effectors ROCK1 and DIA1. Actin meshwork rearrangement regulates ciliary lengthening by controlling the access of transport vesicles to the centrosomal zone. In the absence of Cav1α (right), RhoA activation is impaired and less apical actin polymerizes, allowing the arrival of more material at the centrosome that is used to assemble longer cilia.

Article Snippet: The sources of the antibodies to the different markers were as follows: Cav1 (mouse mAb IgG1, used at 1/2,000 for immunoblot analysis; 610406; and the rabbit polyclonal antibody used at 1/200 for immunofluorescence analysis; 610059) and DIA1 (mouse mAb IgG1, used at 1/1000 for immunoblotting; clone51/mDia1; 610484), were from BD Transduction Laboratories. γ-tubulin (mouse mAb IgG1, used at 1/500 for immunofluorescence analysis; clone GTU-88, T3559) and acetylated tubulin (mouse mAb IgG2b, used at 1/500 for immunofluorescence analysis; clone 6-11B-1; T7451) were obtained from Sigma-Aldrich.

Techniques: Activity Assay, Activation Assay

Journal: Journal of Biological Chemistry

Article Title: Nonredundant roles of DIAPHs in primary ciliogenesis

doi: 10.1016/j.jbc.2021.100680

Figure Lengend Snippet: Figure 1. DIAPH3 regulates ciliogenesis and cilia length. A–C, impairment of ciliation and cilia length by DIAPH2 depletion cannot be rescued by other isoforms. hTERT-RPE1 cells were transfected with siCtr or siDIAPH2#1 for 48 h, and cells were rescued using wild-type constructs of DIAPH1-myc, DIAPH2-myc, and DIAPH3-myc. A, western blots of samples blotted for human DIAPH2, myc, and GAPDH. Quantifications of (B) ciliation and (C) cilia length are shown. Error bars in B represent ± SD of three independent experiments; n = 50 or more each. Graph of cilia length measurement (C) is displayed in box-and-whisker plot, where the upper and lower quartiles are indicated as the ends of the box, the median is marked by a horizontal line inside the box, mean is marked by a cross sign inside the box, and whiskers are two lines outside the box that extend to the highest and lowest observations, while dots outside this range represent outliers. Two-tailed t-test analysis was done to compare control siRNA to all samples or siDIAPH1 to rescue samples, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. D–F, reduction of ciliation and cilia length by DIAPH3 depletion cannot be rescued by other isoforms. hTERT-RPE1 cells were transfected with siCtr or siDIAPH3#1, and cells were rescued using wild-type constructs of DIAPH1-myc, DIAPH2-myc, or DIAPH3-myc. D, western blots of rescued cells blotted for human DIAPH3, myc, and GAPDH. E and F, quantification of ciliation and cilia length in hTERT-RPE1 cells treated with siRNA for 48 h, respectively. Error bars in E represent ±SD of three independent experiments; n = 50 or more each. Two-tailed t-test analysis was done to compare control siRNA to all samples or siDIAPH1 to rescue samples, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. Graph of cilia length measurement is displayed in a box-and-whisker plot as in C. G and H, DIAPH2 and DIAPH3 localize at the base of cilia. hTERT-RPE1 cells were grown to confluence and serum starved for 24 h. Cells were stained with Actub (axonemal marker) and centrin (CNT) for cilia basal marker. Cells were costained for (G) DIAPH2 and CNT or DIAPH2 and Actub. H, cells were costained for DIAPH3 and CNT or DIAPH3 and Actub. Scale bars in G and H are 10 μm in the left panels and 1 μm in the right-side panels.

Article Snippet: BBS1 (21118-1-AP), Arl13B (1711-1-AP), DIAPH3 (14342-1-AP), IFT20 (13615-1- AP), Rab11 (20220-1-AP), and IFT88 (139671-1-AP) were obtained from Proteintech (Rosemont, Illinois, United States).

Techniques: Transfection, Construct, Western Blot, Whisker Assay, Two Tailed Test, Control, Staining, Marker

Journal: Journal of Biological Chemistry

Article Title: Nonredundant roles of DIAPHs in primary ciliogenesis

doi: 10.1016/j.jbc.2021.100680

Figure Lengend Snippet: Figure 2. Decreased trafficking to primary cilia upon depletion of DIAPH2 and DIAPH3. A–D, depletion of DIAPH2 or DIAPH3 decreased trafficking to cilia. A and B, hTERT-RPE1 cells were transfected with siCtr, siDIAPH3, or siDIAPH3 plus the rescue construct myc-DIAPH3 for 48 h. Later cells were fixed and stained for (A) Arl13B and PC1 or (B) acetylated tubulin, Eb3 (as basal body marker) and IFT88. Scale bars are 5.00 μm for the represented images. C, depletion of DIAPH2 reduced in the level of IFT88, PC1, and tubulin acetylation with no change in Arl13B in primary cilia. Box-and-whisker plots (as defined for Fig. 1C) are used to display average fluorescence intensity of Arl13B, PC1, acetylated tubulin, and IFT88 upon DIAPH2 depletion and rescue. Fluorescence intensity was measured as the average intensity over the entire cilium (see Fig. S3E) and therefore normalized for length. The rescue for Arl13B was not done (nd). D, reduction of IFT88 and tubulin acetylation levels was detected upon depletion of DIAPH3. Average fluorescence intensity was measured to determine protein levels of Arl13B, PC1, acetylated tubulin, and IFT88 upon DIAPH3 depletion and rescue. The rescue for Arl13B and PC1 was not done (nd). Box-and-whisker plots are used to display average fluorescence intensity. Three independent experiments; n = 50 cells each. Two-tailed t-test analysis was done to compare control siRNA to all protein depleted samples and as well compare siDIAPH2 or siDIAPH3 to rescue samples, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.

Article Snippet: BBS1 (21118-1-AP), Arl13B (1711-1-AP), DIAPH3 (14342-1-AP), IFT20 (13615-1- AP), Rab11 (20220-1-AP), and IFT88 (139671-1-AP) were obtained from Proteintech (Rosemont, Illinois, United States).

Techniques: Transfection, Construct, Staining, Marker, Whisker Assay, Fluorescence, Two Tailed Test, Control

Journal: Journal of Biological Chemistry

Article Title: Nonredundant roles of DIAPHs in primary ciliogenesis

doi: 10.1016/j.jbc.2021.100680

Figure Lengend Snippet: Figure 3. Targeting DIAPH3 to the ciliary base regulates cilia length and morphology. A–F, targeting DIAPH2 and DIAPH3 to the ciliary base via centrin or PACT causes elongation and bulb formation at the tip of the cilia. A, fusion constructs were made between GFP-centrin (CNTN1) and the indicated proteins (DIAPH2 and DIAPH3). Cells were transfected with GFP-CNTN1 (control), GFP-CNTN1-DIAPH2, or GFP-CNTN1-DIAPH3 and blotted for GFP to detect protein size and to ensure integrity of the fusion proteins. B, constructs were transfected into hTERT-RPE1 cells, which were then serum starved for 24 h to induce ciliogenesis, fixed, and stained for acetylated tubulin (cilia marker) and GFP (to detect transfected cells). Cilia are shown in boxes (left panels) and the boxed areas are magnified (right panels). Scale bars are 8 μm in the left panel and 1 μm in the right-side panels. Arrowheads point toward the bulbs at tips of cilia. C, quantification of bulbed versus nonbulbed cilia. Measurement of cilia length for GFP-CNTN1, GFP-CNTN1-DIAPH2, or GFP-CNTN1-DIAPH3 samples was presented in Box–Whisker graphs (as in Fig. 1C). D, targeting DIAPH3 to the ciliary base via PACT causes elongation and formation of bulbs at the tip of the cilia. Fusion constructs were made between GFP-PACT (centrosomal targeting domain) and DIAPH3. hTERT-RPE1 cells were transfected with GFP-PACT or GFP-PACT-DIAPH3 and serum starved for 24 h and fixed and stained for GFP and acetylated tubulin. Scale bars are 1 μm for each image. E–F, hTERT-RPE1 cells were transfected with CNTN1-GFP (control), GFP-CNTN1-DIAPH1, GFP-CNTN1-DIAPH3, GFP-PACT (control), GFP-PACT-DIAPH1, or GFP-PACT-DIAPH3. Cells were serum starved for 24 h to induce ciliogenesis. Quantification for ciliated cells with bulb (E) and length of bulbed versus nonbulbed cilia (F) was conducted. Error bars in E represent ± SD of three independent experiments; n = 50 cells each, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. Two- tailed t-test analysis was done to compare CNTN1-GFP control to the samples with GFP-CNTN1, while GFP-PACT control was compared with samples with GFP-PACT. Box and whisker plots (F) were performed as in Fig. 1C).

Article Snippet: BBS1 (21118-1-AP), Arl13B (1711-1-AP), DIAPH3 (14342-1-AP), IFT20 (13615-1- AP), Rab11 (20220-1-AP), and IFT88 (139671-1-AP) were obtained from Proteintech (Rosemont, Illinois, United States).

Techniques: Construct, Transfection, Control, Staining, Marker, Whisker Assay, Two Tailed Test

Journal: Journal of Biological Chemistry

Article Title: Nonredundant roles of DIAPHs in primary ciliogenesis

doi: 10.1016/j.jbc.2021.100680

Figure Lengend Snippet: Figure 4. Trafficking of post-Golgi and endosomal vesicles is increased upon targeting DIAPH3 to the base of cilia. A–G, targeting DIAPH3 to the base of cilia increased cargo trafficking. hTERT-RPE1 cells were transfected with GFP-CNTN1 or GFP-CNTN1-DIAPH3, and after 24 h of serum starvation, cells were fixed and stained for acetylated tubulin, GFP and (A) Arf4, (B) IFT20, (C) Rab11, (D) BBS1, or (E) GFP. Scale bars are 3 μm for each image. F, quantifications of the total protein fluorescence intensity scan at the base of cilia was conducted for Arf4, IFT20, Rab11, BBS1, or GFP. G, DIAPH3 targeted to the ciliary base via PACT increases the recycling endosome (Rab11) protein level at the base of the cilia. Cells were transfected with GFP-PACT or GFP-PACT-DIAPH3 and after 24-h serum starvation, were fixed and stained for acetylated tubulin, GFP, and Rab11. Quantification of the Rab11 fluorescence protein intensity scan was conducted. For panels F and G, box and whisker plots (as defined in Fig. 1C) are shown. H, targeting DIAPH3 to the base of cilia has no effect on protein levels of Arf4 or IFT20. Cells were transfected with GFP-CNTN1 (control) or GFP-CNTN1-DIAPH3, and samples were western blotted for GFP, GAPDH, Arf4, and IF20. Three independent experiments were conducted with n = 50 cells each, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. Two-tailed t-test analysis was done to compare GFP-CNTN1 control with GFP-CNTN1-DIAPH3.

Article Snippet: BBS1 (21118-1-AP), Arl13B (1711-1-AP), DIAPH3 (14342-1-AP), IFT20 (13615-1- AP), Rab11 (20220-1-AP), and IFT88 (139671-1-AP) were obtained from Proteintech (Rosemont, Illinois, United States).

Techniques: Transfection, Staining, Whisker Assay, Control, Western Blot, Two Tailed Test

Journal: Journal of Biological Chemistry

Article Title: Nonredundant roles of DIAPHs in primary ciliogenesis

doi: 10.1016/j.jbc.2021.100680

Figure Lengend Snippet: Figure 5. Microtubule function of DIAPH3 regulates ciliogenesis and cilia length. A–C, disruption of microtubule but not actin functions of DIAPH3 negatively impacts ciliogenesis and cilia length. hTERT-RPE1 cells were transfected with siCtr or siDIAPH3, and cells were rescued using wild-type constructs of GFP-DIAPH3WT (wildtype), GFP-DIAPH3ACT (mutation preventing actin polymerization), GFP-DIAPH3MT (mutation preventing microtubule stabilization), GFP-CETN1-DIAPH3, or GFP-CETN1. A, western blots of samples were blotted for human-specific anti-DIAPH3, GFP, and GAPDH. Quantifications for (B) ciliation and (C) cilia length were conducted. D, reduction of IFT20, Rab11, and IFT88 levels at the cilia following DIAPH3 depletion cannot be rescued by DIAPH3MT. Cells were starved for 24 h, fixed, and stained for acetylated tubulin, GFP, and IFT20 or Rab11 or IFT88. For panels C and D, box and whisker plots (as defined in Fig. 1C) are shown. Error bars represent ± SD of three independent experiments; n = 50 each, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. Two-tailed t-test analysis was done to compare siCtr control with all samples or to compare siDIAPH3 with all samples. For panel C, one-way ANOVA statistical testing was also conducted on the all samples excluding the siCtr (p = 0.0001) or including siCtr (p = 0.000004) indicating that a significant difference was detected among populations.

Article Snippet: BBS1 (21118-1-AP), Arl13B (1711-1-AP), DIAPH3 (14342-1-AP), IFT20 (13615-1- AP), Rab11 (20220-1-AP), and IFT88 (139671-1-AP) were obtained from Proteintech (Rosemont, Illinois, United States).

Techniques: Disruption, Transfection, Construct, Mutagenesis, Western Blot, Staining, Whisker Assay, Two Tailed Test, Control